June 25, 1999 

Introduction

Michael's Great Smokies lab reports of 8.7.98, 10.19.98, and 2.8.99 document
intestinal colonization by Klebsiella pneumoniae (Kp) and hemolytic Eschirichia
coli (hEc), which can translocate from gastrointestinal tissue into mesenteric
lymph nodes, peritoneum, and other tissues, including the brain. Whether or not
such translocation has occurred in Michael remains to be determined.

[a]  Serology of Kp-antibodies is well studied in patients inflammatory bowel
disease and ankylosing spondylitis (1a-j). These articles, including one with
Andy Wakefield as a co-author (1a), ought provide helpful context for
interpreting additional Kp data I recommend be obtained for Michael.
Furthermore, his parents might consult with Andy or other authors in the several
Kp-antibodies references, for these researchers have established assays
regarding Kp variants and ought be considered as a primary resource for
interpreting and treating Michael's persistent Kp colonization and possible
spread (as of late June 1999) into other tissues. 

[b] Kp is often a nosocomial infection (2a) and occasionally co-occurs with E.
coli in pathogolic states (2b); but the level and duration of Michael's
colonization indicate that his Kp presence is atypical. I wonder whether or not
he had acquired an unusually high colonization prior to the MMR, which would
have increased intestinal and blood-brain-barrier permeabilities (3a-f) and
augmented tendencies towards bacterial translocation. However, the possibility
of another, co-occurring infection (eg, asymptomatic neonatal CMV) cannot be
ruled out. 

[c]  Generally, Kp does not appear within the biliary system (4a), but the
possibility of Michael having a liver atypicality is suggested by the elevated
copper in early tests and the variability copper data (4b) that *may* reflect
treatments or supplements and alterations in immune and biliary status.

[d]  Although starting Dr. Kane's fatty acid and electrolyte corrections
(4.19.99) would not have affected prior copper levels, supplements that alter
lipid profiles modulate immunological processes, noting that 
     "The leukotrienes are potent lipid mediators of inflammation formed
     by the 5-lipoxygenase-catalyzed oxidation of arachidonic acid." and
     "5-Lipoxygenase reaction products modulate alveolar macrophage
     phagocytosis of Klebsiella pneumoniae." (5a) 
Thus, when his mother reports that Dr. Kane's protocol has been among the most
helpful (along with FGF2), the possibility exists that fatty acid corrections
have been improving Michael's immune defences against systemic or peritoneal
Klebsiella pneumoniae. Reactions to Kp LPS have been studied (eg, 5b).

[e]  Various immune impairments can augment Kp and hEc growth and expansion (6a-
c). Early intestinal exposure to a given pathogen can have that effect via CD5+
immune cell development (6a), and "Certain gram-negative infections have been
shown to induce T-helper-(Th)2 type CD4+ T-cell differentiation, which
correlates with impaired elimination of infection and death.", however, other
early-exposure mechanisms affect weakened immunity against Ec and Kp (6b); and
"Preexposure to hemolytic E coli and other Enterobacteriaceae alters responses
to intra-abdominal infection." (6c); and whereas E. coli does not impair
colonization by Kp (6e), hemolytic E. coli augments the tendency of Klebsiella
pneumoniae to translocate (6d).

[f] Mannose-like molecules are natural parts of Kp cell structure and
participate in macrophage recognition of Kp (7). Astrocytes have mannose
receptors, and the number of these receptors increases in the presence of Th2-
type cytokines patterns, and astrocyte activation is associated with neuronal
dysfunction and even neuronal death (8a-8f). Therefore, circulating Kp fragments
from translocation of intestinal Kp and/or from peritoneal Kp would appear to
have the ability to affect astrocytes located near the blood brain barrier,
thereby affecting associated neurons and neuronal circuits, especially in bbb
regions in which infection-induced vascular atypicality has developed, as
possibility reflected in his MRI results.

[g]  Kp is a T-cell-independent polyclonal activator of immunoglobulin secretion
and and can increase the number of cells in regional lymph nodes (9a-9k; 9l).
Thus, an enduring high level of Kp fragments *may* account for some of Michael's
mildly elevated titres (eg, pertussis and tetanus), but may or may not account
for his various anti-measles elevations (???).

[h]  Classical and alternative complement pathways participate in responses
against Kp, but strain differences can diminish complement's effectivness (10a-
b), and this may be evaluated in Michael if his Kp strain (or strains) is/are
evaluated; and at least one antibiotic has been shown to increase complement
binding to Klebsiella pneumoniae (10c). Futhermore, in certain Kp strains
mannose-related molecules seem to have anti-complement effects and this
corresponds to polyclonal activation (10d).
          "The antibody-independent binding of C1q to serum-sensitive
          strains was independent of the presence of capsular
          polysaccharide, while strains possessing lipopolysaccharide
          O antigen bind less C1q and are resistant to
          complement-mediated killing." (10a)


Teresa Binstock
Researcher in Developmental and Behavioral Neuroanatomy
June 25, 1999

 Summary Why Klebsiella pneumonia and hemolytic E. coli have come to demand much attention in this report. (Kp, hEc)

 Translocation of Kp and hEc

 Peritoneal and abdominal Kp and hEc

 Neurologic aspects of Kp and hEc

 Miscellany regarding Kp and hEc

 Additional tests

 Michael's index page

     A series of autism-spectrum research monographs is available by
     links on a web page: http://www.jorsm.com/~binstock/index.htm

1a. Br J Rheumatol 1998 May;37(5):525-31 
Characterization of the humoral immune response to Klebsiella species in
inflammatory bowel disease and ankylosing spondylitis.
Tiwana H, Walmsley RS, Wilson C, Yiannakou JY, Ciclitira PJ, Wakefield AJ,
Ebringer A
Division of Life Sciences, Infection and Immunity Group, King's College,
London.

This study was carried out to characterize the antibody class response by ELISA
to seven Klebsiella pneumoniae serotypes (K2, K3, K17, K21, K26, K36, K50) in
five different groups, 40 HLA-B27-positive ankylosing spondylitis (AS)
patients, 46 patients with Crohn's disease (CD), 38 patients with ulcerative
colitis (UC), 50 patients with active anti-endomysial antibody-positive coeliac
disease and 40 healthy controls, using whole bacteria and capsular
polysaccharide. IgG antibody levels were significantly elevated in AS patients
to K17, K36, K50; IgA to K2, K3, K21, K26, K36 and K50; and IgM to serotype K21
when compared to normal controls. Furthermore, IgG antibody levels were
significantly elevated in CD patients to K2, K17, K21, K26, K36 and K50; IgA to
K2, K3, K21, K26, K36 and K50; and IgM to K2, K3, K17, K21 and K50. Increased
IgG antibody levels in the UC group were limited only to K17, K36 and K50. No
antibody class was increased to any of the K. pneumoniae serotypes in the
coeliac disease group. The immune responses in AS patients also involve
Klebsiella bacteria having capsular serotypes other than K26, K36 and K50. The
similarity in the immune responses between CD and AS groups suggests that many
AS patients may have occult bowel inflammation.
PMID: 9651080, UI: 98313100

1b. Ann Rheum Dis 1997 Jul;56(7):421-5 
Enhanced jejunal production of antibodies to Klebsiella and other
Enterobacteria in patients with ankylosing spondylitis and rheumatoid
arthritis.
Maki-Ikola O, Hallgren R, Kanerud L, Feltelius N, Knutsson L, Granfors K
National Public Health Institute, Department in Turku, Finland.

OBJECTIVE: To measure gut immunity directly in jejunal fluid in patients with
ankylosing spondylitis (AS) and rheumatoid arthritis (RA). METHODS: Antibodies
against three different Enterobacterias were measured in jejunal perfusion
fluids (collected by a double balloon perfusion device) of 19 patients with AS,
14 patients with RA, and 22 healthy controls using enzyme linked immunosorbent
assay. RESULTS: The AS patients had significantly increased jejunal fluid
concentrations of IgM, IgG, and IgA class antibodies against Klebsiella
pneumoniae, and IgM and IgA class antibodies against Escherichia coli and
Proteus mirabilis compared with healthy controls. When compared with the
patients with RA, the AS patients had higher concentrations of IgA and IgG
class antibodies only against K pneumoniae. The RA patients had higher IgM
class antibody concentrations against all three studied Enterobacterias, when
compared with the healthy controls, suggesting an enhanced mucosal immune
response in these patients. A three month treatment with sulphasalazine did not
decrease enterobacterial antibody concentrations in the 10 patients with AS.
CONCLUSION: There is strong direct evidence for an abnormal mucosal humoral
immune response particularly to K pneumoniae in patients with AS.
PMID: 9486004, UI: 98146958

1c. Ann Rheum Dis 1997 Mar;56(3):180-3 
Association of gut inflammation with increased serum IgA class Klebsiella
antibody concentrations in patients with axial ankylosing spondylitis (AS):
implication for different aetiopathogenetic mechanisms for axial and peripheral
AS?
Maki-Ikola O, Leirisalo-Repo M, Turunen U, Granfors K
National Public Health Institute, Department in Turku, Finland.

OBJECTIVES: A role for Klebsiella pneumoniae in ankylosing spondylitis (AS) has
been suggested because faecal carriage of Klebsiella and serum Klebsiella
specific antibodies may be increased in this disease. This study has extended
the earlier findings by comparing Klebsiella specific serum IgA class
antibodies with inflammatory changes in the gut. METHODS: IgA antibodies to K
pneumoniae, Escherichia coli, and Proteus mirabilis in serum samples of 25
patients with AS, of eight control patients, and of 100 healthy blood donors
were measured by enzyme immunoassay. Gut inflammation of the patients was
studied by ileocolonoscopy. RESULTS: Increased IgA antibody concentrations to
K pneumoniae associated with gut inflammation in patients with axial form of AS.
Such association was not seen in patients with peripheral form of AS.
CONCLUSIONS: These findings may provide further evidence for the role of K
pneumoniae in the pathogenesis of AS. However, at least some of the patients
with axial AS without gut inflammation, as well as patients with peripheral AS
did not have increased K pneumoniae antibody concentrations, which may be
regarded as evidence against the direct role of K pneumoniae in the
pathogenesis. The aetiopathogenetic mechanisms in the axial and peripheral form
of AS may be different.
PMID: 9135221, UI: 97280913

1d. Rheumatol Int 1997;17(1):11-6 
Antibody responses to gut bacteria in ankylosing spondylitis, rheumatoid
arthritis, Crohn's disease and ulcerative colitis.
Tiwana H, Wilson C, Walmsley RS, Wakefield AJ, Smith MS, Cox NL, Hudson MJ,
Ebringer A
Division of Life Sciences, King's College, London, UK.

Specific immunoreactive anti-Klebsiella antibodies are found in patients with
ankylosing spondylitis (AS), a significant proportion of whom have occult
inflammatory bowel disease. Molecular mimicry between Klebsiella or other
bacterial antigens and HLA-B27 has been suggested in the pathogenesis of AS.
The specificity of increased immunoreactivity against Klebsiella remains to be
assessed against the abundant anaerobic bacterial flora, present either in
healthy controls or in patients with ulcerative colitis (UC) and Crohn's
disease (CD). Total immunoglobulin (Ig; IgG, IgA, IgM) immunoreactivity was
measured by ELISA against Klebsiella pneumoniae, Proteus mirabilis, Escherichia
coli and ten anaerobic isolates of the predominant normal bowel flora in 35
patients with active AS, 60 patients with inflammatory bowel disease (30 CD, 30
UC), 60 patients with active rheumatoid arthritis (RA) and 60 healthy controls.
Ig immunoreactivity to K. pneumoniae was significantly elevated in AS (P <
0.001), CD (P < 0.001) and UC (P < 0.001) patients compared with RA patients
and healthy controls. Furthermore, Ig immunoreactivity to P. mirabilis was
significantly elevated only in RA patients, compared with the other
inflammatory groups (P < 0.001) and controls (P < 0.001). There was no
significant antibody response against E. coli or the ten obligate anaerobes in
any of the test groups. The data suggested an increased immune response to
Klebsiella in patients with AS, UC, CD and to Proteus in patients with RA. The
specificity of these responses in some patients supported a possible role for
enteric Klebsiella in the pathogenesis of AS and Proteus in RA. The role of
Klebsiella in inflammatory bowel disease requires further study.
PMID: 9194209, UI: 97337469

1e. Br J Rheumatol 1995 May;34(5):413-7 
Antibodies to Klebsiella pneumoniae, Escherichia coli and Proteus mirabilis in
the sera of patients with axial and peripheral form of ankylosing spondylitis.
Maki-Ikola O, Nissila M, Lehtinen K, Leirisalo-Repo M, Toivanen P, Granfors K
National Public Health Institute, Department in Turku, Finland.

IgM, IgG and IgA class serum antibodies against the whole Klebsiella
pneumoniae, Escherichia coli and Proteus mirabilis bacteria, as well as against
K. pneumoniae and E. coli lipopolysaccharides (LPSs) were studied earlier in
two separate patient populations of 99 and 85 patients with ankylosing
spondylitis (AS) and in 102 healthy blood donors by enzyme immunoassay. In this
study the patients were divided into groups according to the presence or
absence of peripheral arthritis. The patients with peripheral type AS had
increased levels of IgM and IgA class antibodies against K. pneumoniae, whereas
the patients with axial type AS had increased levels of IgG and IgA class
antibodies to K. pneumoniae, as well as IgA class antibodies against E. coli
and P. mirabilis bacteria. Sulphasalazine treatment decreased the IgM and IgA
class antibodies in peripheral AS and IgA class antibodies in axial AS against
K. pneumoniae LPS. The antibody levels were also decreased against E. coli and
P. mirabilis bacteria in the sera of patients with axial AS. The immunological
findings in patients with peripheral and axial form of AS were different from
each other and thus may reflect different aetiopathogenetic mechanisms for
these two types of AS.
Publication Types:  Clinical trial
PMID: 7788168, UI: 95307910

1f. Br Med J (Clin Res Ed) 1988 May 21;296(6634):1432-4 
Raised titres of anti-klebsiella IgA in ankylosing spondylitis, rheumatoid
arthritis, and inflammatory bowel disease.
Cooper R, Fraser SM, Sturrock RD, Gemmell CG
Department of Bacteriology, Glasgow Royal Infirmary.

Serum titres of IgA are raised in ankylosing spondylitis and increased titres
of antibodies to klebsiella have also been reported. The humoral response was
investigated in ankylosing spondylitis and other inflammatory disorders. IgA
antibodies to klebsiella pneumoniae K43 were measured in patients with
ankylosing spondylitis, Crohn's disease, ulcerative colitis, and rheumatoid
arthritis and in controls. Significantly raised median titres of
anti-klebsiella IgA, measured as optical density at 405 nm with an enzyme
linked immunosorbent assay (ELISA), were seen among the patients with
ankylosing spondylitis (0.7), Crohn's disease (0.8), rheumatoid arthritis
(0.6), and ulcerative colitis (0.8) compared with controls (0.4). Activity of
disease in ankylosing spondylitis and titres of anti-klebsiella IgA were not
correlated. In contrast, titres of anti-klebsiella IgM were significantly lower
in patients with ankylosing spondylitis and ulcerative colitis. The increase in
the titres of anti-klebsiella IgA may be due to increased permeability of the
gut to bacterial antigens, leading to an increased IgA response in the gut
mucosa and permitting the release of IgA into the circulation. As the increased
antibody titres were seen in Crohn's disease and rheumatoid arthritis as well
as in ankylosing spondylitis the response may be nonspecific, occurring because
of possible underlying inflammatory bowel disease in these conditions.
PMID: 3132277, UI: 88241301

1g. Br J Rheumatol 1998 Dec;37(12):1299-302 
IgA class serum antibodies against three different Klebsiella serotypes in
ankylosing spondylitis.
Maki-Ikola O, Nissila M, Lehtinen K, Granfors K
National Public Health Institute, Department in Turku, Finland.

OBJECTIVE: To investigate the possible predominance of certain Klebsiella
pneumoniae capsular types in the pathogenesis of ankylosing spondylitis (AS).
METHODS: The prevalence of IgA class antibodies against three different K.
pneumoniae strains (with capsular types 21, 30 and 43) was studied in the sera
of 177 patients with AS and of 100 healthy blood donors using an enzyme-linked
immunosorbent assay. RESULTS: The median Klebsiella-specific antibody levels
were always higher in patients than in controls regardless of the serotype used
as antigen. When the prevalence of increased antibody levels was compared
between the groups, it was highest against the strain with capsular type 30,
whereas against strains 21 and 43 it was similar among patients and controls.
CONCLUSIONS: A broad range of Klebsiella serotypes may be involved in the
pathogenesis of AS. Thus, it is important to take the different Klebsiella
serotypes into particular account in these studies.
PMID: 9973153, UI: 99137438

1h. J Rheumatol 1998 Sep;25(9):1756-64 
Antibody response to Klebsiella pneumoniae 60 kDa protein in familial and
sporadic ankylosing spondylitis: role of HLA-B27 and characterization as a
GroEL-like protein.
Cancino-Diaz ME, Perez-Salazar JE, Dominguez-Lopez L, Escobar-Gutierrez A,
Granados-Arreola J, Jimenez-Zamudio L, Burgos-Vargas R, Garcia-Latorre E
Departamento de Inmunologia, Escuela Nacional de Ciencias Biologicas, Instituto
Politecnico Nacional, Hospital General de Mexico SSA, DF, Mexico.

OBJECTIVE: To study the antibody response of HLA-B27+ patients with ankylosing
spondylitis (AS) and their first degree relatives to the 60 kDa protein of
Klebsiella pneumoniae and to characterize this protein. METHODS: Sera from 84
individuals were analyzed by ELISA to determine the titer of antibodies against
the 60 kDa protein of K. pneumoniae. Subjects were divided into 3 categories:
Group 1: 44 HLA-B27+ AS related individuals (35 patients, 9 healthy controls);
Group 2: 28 healthy B27- AS related individuals; and Group 3: 12 healthy B27-
non-AS related subjects. The 60 kDa protein of K. pneumoniae was induced at 45
degrees C and purified by electroelution from sodium dodecyl
sulfate-polyacrylamide gel electrophoresis. It was characterized as a
GroEL-like heat shock protein (HSP). The recognition of GroEL-like protein was
confirmed by immunoblot of 2 dimension electrophoresis. The response to
GroEL-like protein from other bacteria and the response to lipopolysaccharide
(LPS) was also analyzed by immunoblot. RESULTS: HLA-B27+ individuals (Group 1),
independent of their disease status, showed a significant higher response to
the 60 kDa protein of K. pneumoniae than HLA-B27- subjects from Groups 2 and 3
(p < 0.0001). This protein was characterized as a HSP of the GroEL family and
designated HSP60Kp. The GroEL of other enterobacteria as well as that of
Mycobacterium leprae were recognized by HLA-B27+ individuals by immunoblot,
whereas HLA-B27- individuals did not. LPS was not recognized by HLA-B27
positive or negative subjects. CONCLUSION: These findings suggest a
relationship between HLA-B27 and the response to a GroEL-like protein that
could have implications in AS.
PMID: 9733457, UI: 98402293

1i. Microb Pathog 1998 Jul;25(1):23-32 
Cloning and sequencing of the gene that codes for the Klebsiella pneumoniae
GroEL-like protein associated with ankylosing spondylitis.
Cancino-Diaz M, Curiel-Quesada E, Garcia-Latorre E, Jimenez-Zamudio L
Departamento de Immunologia, Escuela Nacional de Ciencias Biologicas, Instituto
Politecnico Nacional, Mexico D.F., Mexico.

Ankylosing spondylitis (AS) is a chronic inflammatory disease of the sacroiliac
joints and vertebral column of unknown aetiology, but strongly related to the
presence of the HLA-B27 antigen. The participation of bacterial infections as
triggering factors have also been suggested. We have associated the 60 kDa heat
shock protein of Klebsiella pneumoniae (HSP60Kp) with AS since we have
previously demonstrated that most of the patients have IgG antibodies and
active T cells that recognize preferentially this protein, but we have not yet
identified the epitopes involved in the recognition. In order to know the amino
acid sequence of HSP60Kp, and to be able to analyse in the future the relevant
epitopes; we amplified by PCR and cloned the gene coding for this protein into
the SmaI site of pUC19. The nucleotide sequence of the gene was obtained by the
Sanger method using both manual and automatic techniques. Amino acid sequence
of the HSP60Kp was deduced by translating the nucleotide sequence of the gene.
The antigenic analysis of this sequence was compared to the antigenic analysis
of the reported sequences of Escherichia coli GroEL and Yersinia enterocolitica
HSP60. Using a software to predict HLA class I motifs, the nonapeptide
(KRGIDKAVL) residues 117-125 of HSP60Kp showed a much higher affinity for
HLA-B27 than the similar nonapeptide of E. coli GroEL and Y. enterocolitica
HSP60. The only difference between the three peptides was in position nine.
This finding could explain the association of AS only with the HSP60 of
Klebsiella pneumoniae. On the other hand, hydrophilicity analysis, which
indicates B cell epitopes, showed three similar strongly antigenic regions in
the three proteins.
PMID: 9705246, UI: 98379446

1j. J Rheumatol 1998 Apr;25(4):743-7 
Antibodies to Klebsiella pneumoniae in Dutch patients with ankylosing
spondylitis and acute anterior uveitis and to Proteus mirabilis in rheumatoid
arthritis.
Blankenberg-Sprenkels SH, Fielder M, Feltkamp TE, Tiwana H, Wilson C, Ebringer
A
Netherlands Ophthalmic Research Institute, Amsterdam.

OBJECTIVE: To determine whether the association between increased humoral
reactivity against Klebsiella and HLA-B27 associated diseases could be
confirmed in Dutch patients with ankylosing spondylitis (AS) and acute anterior
uveitis (AAU). METHODS: Under coded conditions sera from Dutch patients with
AS, AAU, and rheumatoid arthritis (RA) and from HLA-B27 positive and negative
healthy controls were studied for IgA anti-Klebsiella (K54) and IgG
anti-Proteus antibodies with the indirect immunofluorescence assay on whole
bacteria fixed in suspension with paraformaldehyde. Each group consisted of at
least 17 sera. RESULTS: IgA anti-Klebsiella antibody titers were elevated in AS
and HLA-B27 negative AAU compared to the HLA-B27 positive and negative controls
or patients with active RA (p < 0.001). Furthermore, patients with active RA
had elevated levels of IgG antibodies against P. mirabilis compared to every
other test or control group (p < 0.001). There was no significant difference
between the AS and RA patients in terms of serum C-reactive protein levels,
although these were significantly elevated in both compared to healthy controls
(p < 0.001), suggesting that the antibody elevations were not due to a
nonspecific inflammatory effect. The same sera were blindly tested with
negative results by 2 other centers. The discrepancies are probably the result
of differences in the methods used. CONCLUSION: Our data support the hypothesis
that Klebsiella are involved in the pathogenesis of AS and AAU and that the
same might be true for Proteus in RA.
PMID: 9558179, UI: 98217262


2a. Scand J Infect Dis 1991;23(2):189-94 
Spread of Klebsiella in a neonatal ward.
Hambraeus A, Lagerqvist-Widh A, Zettersten U, Engberg S, Sedin G, Sjoberg L
Department of Clinical Bacteriology, University Hospital, Uppsala, Sweden.

The colonization of infants with Klebsiella pneumoniae was prospectively
studied. Samples were taken from nose, throat, umbilicus and rectum on the day
of arrival and thereafter once a week. Phage typing was performed the first
time K. pneumoniae was found at any of these sites. Settle plates were exposed
in the incubators and in the patient rooms 5 h/day. The study lasted for 32
weeks. The first 15 weeks was a control period with no information to the
staff, the following 4 weeks was a period of intervention and education and the
last 13 weeks was a second control period. In all, 603 infants were
investigated. The number of infants nursed per week and severity of their
disease was comparable in the 3 periods. The colonization rates were 65, 34 and
58%, respectively. The acquisition of new strains was 1.4 per infant in the
first and last periods, but only 0.4 in the period of intervention. Thus,
colonization rates decreased only during the period of continuous education in
hygiene.
PMID: 1853167, UI: 91305893

2b. Acta Paediatr Scand 1982 May;71(3):457-65 
Colonization of the upper jejunum by enteropathogenic and enterotoxigenic
Escherichia coli in paediatric diarrhoea.
Stintzing G, Mollby R

Aspirated fluid from the upper jejunum was obtained by intubation of 27
children with diarrhoea and 7 control children without diarrhoea. The aspirated
jejunal fluid was analysed for total counts of viable aerobic and anaerobic
microorganisms. Gram-negative aerobic bacteria were typed biochemically and
analysed for the production of heat-labile and heat-stable enterotoxin.
Enteropathogenic Escherichia coli (EPEC) strains of serogroups 0111, 055 and
0127 were detected in the stools of nine patients and the respective strains
concomitantly in the upper jejunum (10(3)-10(8) bact/ml) in 7 patients with
diarrhoea. In 6 patients from whom isolates of enterotoxigenic E. coli (ETEC)
were obtained, there were high total viable counts of non-enterotoxigenic
bacteria in the upper jejunum and ETEC were recovered from this location in 3
cases. Enterotoxigenic Klebsiella was recovered from faeces but not from upper
jejunum in one case. Compared to the controls, the total number of bacteria in
the upper jejunum were 100-1000 times higher in patients with enteropathogenic
E. coli (EPEC) or enterotoxigenic E. coli (ETEC). In another 11 cases with
diarrhoea caused by Shigella, rotavirus or of unknown aetiology, the total
viable counts of bacteria were similar to those of the controls. Five patients
with severe EPEC diarrhoea received antibodies for 5 days. The patients
improved clinically, and the EPEC strain had disappeared from the upper jejunum
when they were reexamined. In conclusion, in EPEC diarrhoea a colonization of
the upper jejunum by the causative organism seemed to take place, while in ETEC
diarrhoea, there appeared to be a nonspecific contamination by enteric
bacteria.
PMID: 6753475, UI: 83044547


3. Citations 3a-f were originally presented as part of a brief monograph to the
autism list of St. Johns Univeristy, Posting number 37859, dated 27 Feb 1998,
and entitled "Vaccinations: MMR, IFN-gamma, viral infections".

3a. Pabst HF et al. Kinetics of immunologic responses after primary MMR
vaccination.  Vaccine.  15(1):10-4, 1997 Jan.
  To study the kinetics of humoral as well as cellular immunity to measles and
to test for associated immunosuppression 124 12 month old children were studied
twice, before routine MMR and either 14, 22, 30, or 38 days after vaccination.
Plaque reduction neutralization (PRN) titres were determined at these time
points and lymphocytes were evaluated to identify changes in proportions of
phenotype, their capacity to generate cytokines and to respond to blast
transformation (BT) to measles hemagglutinin (HA), tetanus toxoid and Candida
antigen. The PRN titre and BT to HA plateaued at 30 days and CD8+ and NK cells
increased after immunization. Interleukin 2, 4, and 10 showed no significant
changes. There was mild suppression of BT at 14 and 22 days post-immunization.
Interferon-gamma was the principal cytokine produced after primary measles
immunization, suggesting primary measles immunization induces predominantly a
TH1 type response.

3b. Madara JL.  Stafford J. Interferon-gamma directly affects barrier function
of cultured intestinal epithelial monolayers. Journal of Clinical Investigation.

83(2):724-7, 1989 Feb.
ab:  Although epithelia, which often are in intimate contact with lymphoid
cells, may bear receptors for various cytokines, it is unclear whether cytokines
directly effect epithelial function. We examine the effects of the cytokine
interferon (IFN) on barrier function of cultured monolayers of the T84 human
intestinal epithelial cell line. Gamma IFN, in concentrations and exposures
required to show its other biological effects, directly affects such monolayers.
Monolayer resistance is substantially diminished by gamma IFN. Such effects were
not due to cytotoxicity as judged morphologically and by LDH assays. Solute
fluxes and dual Na+-mannitol flux analysis indicate that the resistance decrease
is due to an effect of gamma IFN on tight junction permeability. The effects of
gamma IFN on monolayer barrier function were not duplicated by the cytokines
interleukin 1, interleukin 2, or tumor necrosis factor. We speculate that such
products of activation of lymphoid cells might influence barrier function of
intestinal, and perhaps other epithelia in disease states.

3c)  Huynh HK.  Dorovini-Zis K. Effects of interferon-gamma on primary cultures
of human brain microvessel endothelial cells.
          American Journal of Pathology.  142(4):1265-78, 1993 Apr.
  Primary cultures of human brain microvessel endothelial cells were used to
study the effects of human recombinant interferon-gamma (IFN-gamma) on cerebral
endothelium in vitro. Incubation of monolayers with various concentrations of
IFN-gamma (10 to 200 U/ml) for 12 to 96 hours induced surface expression of
class II major histocompatibility complex (Ia) antigen in a time- and
concentration-dependent manner. In immunogold-stained cultures, labeling was
observed as early as 12 hours, was maximal after 48 hours, and persisted at
plateau levels in the continuous presence of the cytokine. Expression was
blocked by coincubation with anti-IFN-gamma antibody and was reversed 4 days
following removal of IFN-gamma from the culture media. Endothelial cells treated
with IFN-gamma for 3 to 4 days became spindle-shaped, extensively overlapped,
and frequently formed cellular whorls. These changes did not occur in the
presence of anti-IFN-gamma antibody and reversed upon removal of IFN-gamma from
the media. The morphological alterations were associated with increased
permeability of confluent monolayers to macromolecules as compared with
untreated cultures. The results of these studies indicate that human brain
microvessel endothelial cells respond to in vitro cytokine stimulation by
undergoing profound morphological, functional, and permeability changes. We
conclude that cerebral endothelium may play an important role in the initiation
and regulation of lymphocyte traffic across the blood-brain barrier in
inflammatory disorders of the human central nervous system.

3d. Planchon SM et al. Regulation of intestinal epithelial barrier function by
TGF-beta 1. Evidence for its role in abrogating the effect of a T cell cytokine.

Journal of Immunology.  153(12):5730-9, 1994 Dec 15.
ab: Maintenance of the integrity of the single-cell-thick intestinal epithelium
as an in vivo barrier between environmental Ags and mucosal immunocytes is
pivotal for health. The T cell cytokine IFN-gamma consistently disrupts this
epithelial barrier in vitro...

3e. McRoberts JA.  Riley NE. Modulation of growth factor and cytokine-induced
increases in T84 cell monolayer permeability by media components. Am J of
Physiology.  267(2 Pt 1):C537-43, 1994 Aug.
ab:   Previous studies have shown that insulin, insulin-like growth factors, and
interferon-gamma cause an increase the paracellular permeability across T84
human colonic epithelial cell monolayers... Together, these results strongly
suggest that insulin, insulin-like growth factors, and interferon-gamma increase
the permeability across T84 cell monolayers through a common mechanism that is
modulated by glucose-derived metabolites and oxidative metabolism.

3f. Adams RB et al. IFN-gamma modulation of epithelial barrier function. Time
course, reversibility, and site of cytokine binding. Journal of Immunology. 
150(6):2356-63, 1993 Mar 15.
ab:  The single cell-thick intestinal epithelium forms a crucial barrier between
the host and environment, and is modeled in vitro by a monolayer of polarized,
highly differentiated T84 epithelial cells impermeable to most macromolecules
because of functional intercellular tight junctions.       [Intestinal CMV is
another way to increase permeability.] ...our data demonstrate 1) only the
monolayer's basolateral surface is IFN-gamma responsive, whereas the apical
(microvillous) surface is no; 2) the alteration in electrical resistance of
epithelium is prolonged (5 days), even after a single (24 h) exposure to
IFN-gamma, but nevertheless is reversible; 3) the effect is likely
receptor-ligand mediated, because it can be partially blocked by IFN-gamma
receptor-specific monoclonal Ig; 4) an alteration in tight junction function (a
paracellular pathway) rather than cell necrosis or a transcellular pathway is
responsible for IFN-gamma-induced monolayer dysfunction because permeability to
a 44,000-Da macromolecule (horseradish peroxidase) did not increase, and
intracytoplasmic T84 cell enzymes were not released into the media; and 5) the
biologic phenomenon could not be induced by a species (alpha) of class I IFN,
making IFN-gamma reasonably unique in this regard. Given the proximity;
activation status, and capacity of T lymphocytes for cytokine production in
mucosa, we suggest that IFN-gamma-induced changes in epithelial permeability may
be a major cause of altered intestinal barrier function in vivo.


4. Arch Surg 1977 Aug;112(8):965-7 
Bacteriology of the human biliary tract and the duodenum.
Lou MA, Mandal AK, Alexander JL, Thadepalli H

Using the modern anaerobic transport media and meticulous culture techniques,
74 patients undergoing biliary tract surgery were studied. The biliary system
was found to be sterile in 58 patients (78%). Fifteen patients had 35 isolates
of aerobic and facultative bacteria. The most common ones were Klebsiella,
Enterococcus, and Escherichia coli. The only anaerobe isolated was Clostridium
perfringens. Eight of 17 patients (47%) with acute cholecystitis and five of 49
patients (10%) with chronic cholecystitis, harbored bacteria in the biliary
system. This study suggests that anaerobes are rare in the human biliary
system; therefore, if antibiotic therapy is considered, aerobic coverage should
suffice.
PMID: 195558, UI: 77220892

4b. Micahel's copper data are highly variable. I do not know how typical this
is, especially in young children with known intestinal pathogens capable of
translocating into mesenteric tissues and beyond. 

     blood     6.23.97 high mid-range        MetaMetrix
     hair      9.19.97   mid-range           Great Smokies
     hair      12.1.97   very high           Doctors Data

     blood     2.3.98  low-mid-range         MetaMetrix
     hair      2.19.98   low                 Doctors Data
     blood     4.23.98 above high limit      MetaMetrix
     blood     10.8.98 towards low limit     MetaMetrix
     

5a. Infect Immun 1998 Nov;66(11):5140-6 
5-Lipoxygenase reaction products modulate alveolar macrophage phagocytosis of
Klebsiella pneumoniae.
Mancuso P, Standiford TJ, Marshall T, Peters-Golden M
Division of Pulmonary and Critical Care Medicine, Department of Internal
Medicine, University of Michigan Medical Center, Ann Arbor, Michigan
48109-0642, USA. petersm@medmail.med.umich.edu

The leukotrienes are potent lipid mediators of inflammation formed by the
5-lipoxygenase-catalyzed oxidation of arachidonic acid. Although the effects of
leukotrienes on neutrophil chemotaxis and activation have been established,
their role in modulating innate host defense mechanisms is poorly understood.
In a previous study (M. Bailie, T. Standiford, L. Laichalk, M. Coffey, R.
Strieter, and M. Peters-Golden, J. Immunol. 157:5221-5224, 1996), we used
5-lipoxygenase knockout mice to establish a critical role for endogenous
leukotrienes in pulmonary clearance and alveolar macrophage phagocytosis of
Klebsiella pneumoniae. In the present study, we investigated the role of
specific endogenous leukotrienes in phagocytosis of K. pneumoniae and explored
the possibility that exogenous leukotrienes could restore phagocytosis in
alveolar macrophages with endogenous leukotriene synthesis inhibition and
enhance this process in leukotriene-competent cells. Rat alveolar macrophages
produced leukotriene B4 (LTB4), LTC4, and 5-hydoxyeicosatetraenoic acid
(5-HETE) during the process of phagocytosis, and the inhibition of endogenous
leukotriene synthesis with zileuton and MK-886 dramatically attenuated
phagocytosis. We also observed a reduction in phagocytosis when we treated
alveolar macrophages with antagonists to the plasma membrane receptors for
either LTB4, cysteinyl-leukotrienes, or both. In leukotriene-competent cells,
LTC4 augmented phagocytosis to the greatest extent, followed by 5-HETE and
LTB4. These 5-lipoxygenase reaction products demonstrated similar relative
abilities to reconstitute phagocytosis in zileuton-treated rat alveolar
macrophages and in alveolar macrophages from 5-lipoxygenase knockout mice. We
conclude that endogenous synthesis of all major 5-lipoxygenase reaction
products plays an essential role in phagocytosis. The restorative and
pharmacologic effects of LTC4, LTB4, and 5-HETE may provide a basis for their
exogenous administration as an adjunctive treatment for patients with
gram-negative bacterial pneumonia.
PMID: 9784515, UI: 99003121

5b. J Investig Allergol Clin Immunol 1998 Mar-Apr;8(2):94-7 
Bacterial lipopolysaccharide-induced sulfidoleukotriene release from peripheral
blood leukocytes in patients with asthma and chronic obstructive pulmonary
disease.
Kraus-Filarska M, Malolepszy J, Medrala W, Dobek R
Department of Internal Medicine and Allergology, Wroclaw University of
Medicine, Poland.

Bacterial endotoxins are seen to possess strong proinflammatory activities.
These substances may intensify inflammation in the airways of patients with
chronic obstructive pulmonary disease (COPD) and asthma by facilitating release
of various mediators from different types of cells. Sulfidoleukotrienes (sLT)
cause bronchoconstriction, increase vascular permeability and stimulate mucous
secretion. The aim of our study was to evaluate sLT release from peripheral
blood leukocytes stimulated by Klebsiella pneumoniae lipopolysaccharide (LPS)
and obtained from COPD and asthma patients. Nineteen subjects with mild or
moderate stable bronchial asthma, nine patients with COPD and 10 healthy
controls entered the study. Cellular allergen stimulation test (CAST)-ELISA
test was performed using Buhlmann Laboratories AG kits to determine sLT
production. The differences between atopic (462.57 SD = 215.89 pg/ml) and
nonatopic (474.25 SD = 158.02 pg/ml) asthmatics in comparison to healthy
controls (191.55 SD = 53.2 pg/ml) were statistically significant (p < 0.005)
upon LPS stimulation at the concentration of 10 micrograms/ml. At lower LPS
concentration (1 microgram/ml) the difference was statistically significant
only between nonatopic asthmatics and healthy subjects (p < 0.02). In the COPD
group the sLT production in either LPS concentration was higher than in the
controls but the difference was not significant. We suppose that leukocytes
obtained from asthmatics and COPD patients are more susceptible to LPS than
these cells from healthy individuals. An increased sLT production upon LPS
stimulation during respiratory bacterial infection may intensify inflammation,
bronchoconstriction and increase nonspecific bronchial hyperreactivity.
PMID: 9615302, UI: 98277529

6a. See citations regarding neonatal CD5 immune impairments at
http://www.jorsm.com/~binstock/cd5-iga.htm

6b. Surgery 1998 Aug;124(2):418-28 
Bacterially preexposed T cells impair bacterial elimination by non-Th1/Th2 cell
mechanisms in a model of intra-abdominal infection.
Gleason TG, Sawyer RG, Pruett TL
Surgical Infectious Disease Laboratory, University of Virginia, Charlottesville
22908, USA.

BACKGROUND: Escherichia coli preexposure in mice results in impaired
elimination of subsequent intra-abdominal infections by a CD+4 T cell-dependent
process. Certain gram-negative infections have been shown to induce
T-helper-(Th)2 type CD4+ T-cell differentiation, which correlates with impaired
elimination of infection and death. We hypothesized that E coli preexposure
impairs subsequent bacterial elimination as a consequence of Th2
differentiation and that interleukin-12 (IL-12) treatment could reverse this
differentiation and minimize the effects of E coli preexposure. METHODS: After
preexposure to E coli or other species, BALB/c mice or interferon-gamma
(INF-gamma)-deficient mice, treated with or without IL-12, were given a
standard intra-abdominal infection (E coli, Bacteroides fragilis, and
adjuvant). Cohorts were killed for abscess quantification, in vitro T-cell
proliferative responsiveness, and cytokine secretory profiles. Splenic
lymphocytes preexposed in vivo to other types of bacteria were transferred to
naive mice before intra-abdominal infection to determine whether preexposure,
eliciting the lymphocyte-dependent response, was species specific. RESULTS: E
coli preexposure alone caused no Th1 or Th2 shift; increased the proliferative
responses of T cells; and, in combination with IL-12 therapy, caused markedly
decreased IL-2 and IL-4 responses and an increased IFN-gamma response. IL-12
therapy did not change the response to intra-abdominal infection despite its
ability to cause marked Th1 polarization. IFN-gamma-deficient mice responded to
E coli preexposure no differently than did wild-type mice. Transfer of
lymphocytes preexposed to Pseudomonas aeruginosa, Klebsiella pneumoniae, and
hemolytic E coli but not other types of nosocomial pathogens caused the
development of more abscesses just as transfer of E coli preexposed lymphocytes
had. CONCLUSIONS: CD4+ T cells responsive to E coli preexposure regulate
subsequent intra-abdominal abscess formation by a mechanism not explained by
the Th1/Th2 paradigm. Preexposure to hemolytic E coli and other
Enterobacteriaceae alters responses to intra-abdominal infection.
PMID: 9706167, UI: 98371424

6c. Nat Med 1998 May;4(5):615-8 
Mice lacking neutrophil elastase reveal impaired host defense against gram
negative bacterial sepsis.
Belaaouaj A, McCarthy R, Baumann M, Gao Z, Ley TJ, Abraham SN, Shapiro SD
Respiratory & Critical Care, Department of Medicine, Cell Biology, Washington
University School of Medicine at Barnes-Jewish Hospital, St. Louis, MO 63110,
USA.

Neutrophil elastase (NE) is a potent serine proteinase whose expression is
limited to a narrow window during myeloid development. In neutrophils, NE is
stored in azurophil granules along with other serine proteinases (cathepsin G,
proteinase 3 and azurocidin) at concentrations exceeding 5 mM. As a result of
its capacity to efficiently degrade extracellular matrix, NE has been
implicated in a variety of destructive diseases. Indeed, while much interest
has focused on the pathologic effects of this enzyme, little is known regarding
its normal physiologic function(s). Because previous in vitro data have shown
that NE exhibits antibacterial activity, we investigated the role of NE in host
defense against bacteria. Generating strains of mice deficient in NE (NE-/-) by
targeted mutagenesis, we show that NE-/- mice are more susceptible than their
normal littermates to sepsis and death following intraperitoneal infection with
Gram negative (Klebsiella pneumoniae and Escherichia coli) but not Gram
positive (Staphylococcus aureus) bacteria. Our data indicate that neutrophils
migrate normally to sites of infection in the absence of NE, but that NE is
required for maximal intracellular killing of Gram negative bacteria by
neutrophils.
PMID: 9585238, UI: 98244637

6d. J Med Microbiol 1990 Sep;33(1):17-22 
The role of Escherichia coli haemolysin in the pathogenic synergy of colonic
bacteria in subcutaneous abscess formation in mice.
Ushijima T, Takahashi M, Seto A
Department of Microbiology, Shiga University of Medical Science, Japan.

The growth of nine species of colonic bacteria--Escherichia coli, Enterococcus
faecalis, Bacteroides ovatus, Fusobacterium varium, Clostridium perfringens,
Klebsiella pneumoniae, Proteus vulgaris, Staphylococcus aureus and
Bifidobacterium adolescentis--was examined after concomitant injection to form
experimental subcutaneous abscesses in mice. Injection of a mixture of c. 10(5)
cfu of each of the first five strains (E. coli, Ent. faecalis, B. ovatis, F.
varium and C. perfringens) resulted in abscess formation in all mice tested
when the E. coli strain was haemolytic. E. coli and B. ovatus multiplied and
reached a maximum population of c. 10(8) cfu/abscess. When non-haemolytic E.
coli was used, injection of greater than or equal to 10(7) cfu was required for
abscess formation. The inclusion of partially purified E. coli haemolysin (125
HU50) with c. 10(5) cfu of bacteria including non-haemolytic E. coli resulted
in abscess formation in most mice tested. These results indicate that E. coli
haemolysin is one factor that may potentiate pathogenic synergy among colonic
bacteria especially between E. coli and B. ovatus, during abscess formation.
PMID: 2146395, UI: 91039235

6e. J Antimicrob Chemother 1990 Sep;26(3):411-8 
The contribution of Escherichia coli to microbial colonization resistance.
Vollaard EJ, Clasener HA, Janssen AJ
Department of Pharmacy, Canisius-Wilhelmina Hospital, Nijmegen, The
Netherlands.

The contribution of Escherichia coli to the microbial colonization resistance
(CR) of the bowel was investigated in six healthy volunteers. Esch. coli was
eliminated from faeces by the administration of a low dose (20 mg daily) of
pefloxacin. This did not cause an increase in the faecal concentration of
aerobic Gram-positive cocci or yeasts, nor did it facilitate colonization of
the bowel by a pefloxacin-resistant challenge strain of Klebsiella pneumoniae.
Therefore, Esch. coli does not appear to contribute to the microbial CR. After
ten days of pefloxacin, clindamycin 300 mg was administered twice daily for 18
days. Clindamycin caused a significant increase in the faecal concentration of
enterococci, yeasts and the K. pneumoniae challenge strain, indicating that the
study design was suitable to demonstrate disturbance of microbial CR if it
occurred.
PMID: 2228829, UI: 91035035


7a. Adv Dent Res 1997 Apr;11(1):43-9 
Phagocyte-bacteria interactions.
Keisari Y, Kabha K, Nissimov L, Schlepper-Schafer J, Ofek I
Department of Human Microbiology, Sackler Faculty of Medicine, Tel-Aviv
University, Israel.

Recognition and phagocytosis of micro-organisms in a serum-poor environment
represent innate immunity against many extracellular pathogens. As a paradigm
for such processes, we discuss the recognition of Klebsiella pneumoniae by
alveolar macrophages and monocyte-derived macrophages in the absence of serum.
Macrophages recognize and subsequently kill Klebsiella expressing Man-alpha
2/3-Man or Rha-alpha 2/3-Rha sequences in their capsular polysaccharides by two
mechanisms: (a) recognition of the capsular structures by macrophage mannose
receptors, and (b) opsonization by the lung surfactant protein A (SP-A), which
binds to the capsular polysaccharides of Klebsiella and to SP-A receptors on
the macrophages. Sp-A may also enhance phagocytosis by increasing the activity
of macrophage mannose receptors. We conclude that a specific microbial surface
structure may be a target for recognition by macrophages via several
mechanisms, as exemplified in the case of Klebsiella capsular polysaccharides.
Multiple recognition mechanisms of pathogens by macrophages may be essential to
provide innate immunity to reduce the frequency of infections caused by a
relatively less virulent bacterium in the immuno-compromised host.
Publication Types:  Review  Review, tutorial
PMID: 9524441, UI: 98185127


8a. Glia 1998 Jan 1;25(1):44-55 
Identification and functional characterization of the mannose receptor in
astrocytes.
Burudi EM, Riese S, Stahl PD, Regnier-Vigouroux A
Department of Neurobiology, University of Heidelberg, INF 364, Heidelberg,
Germany.

The immune competence of astrocytes is still ill defined, especially their
endocytic capacity, a prerequisite for efficient antigen presentation. We show
that mannose receptor, a very important conduit for internalization of
infectious agents and self antigens, is functionally expressed in the murine
CNS. By in vitro assays, astrocytes and microglia were shown to be the prime
cells expressing this receptor. Studies on astrocytes demonstrate that its
expression and function are inversely regulated by anti-and pro-inflammatory
compounds. Downregulation of the mannose receptor by IFN-gamma is concomitant
with the induction of the invariant chain, which is also induced by GM-CSF +
IL-4. Mannose receptor-expressing astrocytes may thus act as scavenger not only
in CNS development but also in defense, against soluble and particulate
mannosylated pathogens, presenting fragments thereof at strategic locations in
the CNS. These findings unravel a new and putatively very important role of
astrocytes in innate immunity and possibly development. 
PMID: 9888297, UI: 99103372

8b. Neurotoxicology 1998 Apr;19(2):269-81 
Astrocytes as mediators of immune and inflammatory responses in the CNS.
Aschner M
Department of Physiology and Pharmacology, Bowman Gray School of Medicine of
Wake Forest University, Winston-Salem, NC 27157-1083, USA.

The long-standing view that the brain is isolated from the effects of the
immune system has recently been challenged, with experimental evidence
suggesting that in response to invasion by microorganisms, the CNS can mount
its own defense by resident cells, such as the microglia and astrocytes. Both
cell types produce and secrete a number of cytokines and therefore can
potentially modulate and integrate the communication between hematogenous cells
and resident cells of the CNS. This manuscript will commence with a brief
overview of astrocytic functions in the CNS, and proceed to discuss astrocytic
responses that may regulate CNS inflammation. Specifically, it will address (1)
the function of astrocytes as the antigen presenting cells (APCs) of the CNS,
and (2) the role afforded by astrocyte-derived cytokines, and astrocytic
responses to cytokines secreted elsewhere, in mediating and sustaining immune
responses. Finally, some recent experimental evidence on the possibility that
astroglial impairment by pathogens may contribute to the etiology of neurologic
diseases will be highlighted.

8c. Toxicol Lett 1998 Dec 28;102-103:283-7 
Immune and inflammatory responses in the CNS: modulation by astrocytes.
Aschner M
Department of Physiology and Pharmacology, Wake Forest University School of
Medicine, Winston-Salem, NC 27157-1083, USA. maschner@bgsm.edu

Because the skull bones, the cerebrospinal fluid, the blood-brain barrier
(BBB), and the meninges effectively shield the central nervous system from
other tissues, it was proposed that the brain is an 'immunologically
privileged' organ. However, with recent evidence that in response to invasion
by microorganisms, resident cells, such as astrocytes and microglia can fully
mount an immune response, this long-standing view has been rethought and
revised. Over the last two decades, both astrocytes and microglia have been
shown to secrete numerous cytokines, and, therefore, it is presently widely
accepted that these cells actively participate in an integrative communicative
pathway between resident immune cells of the CNS and those of the periphery.
While clearly implicated in the initiation, maintenance, and suppression of
immune responses, cytokines produced by these cells (e.g. astrocytes and
microglia), as well as the responses of these cells to cytokines produced
elsewhere, has also been shown to propagate CNS damage. Therefore the potential
involvement of these cells in neurodegenerative disorders has been raised and
subjected to intense experimentation. The objective of this synopsis is to
review the role played by astrocytes in the initiation and modulation of immune
responses.


8d. Neurochem Res 1998 May;23(5):759-65 
Neuronal death: is there a role for astrocytes?
Tacconi MT
Istituto di Ricerche Farmacologiche Mario Negri, Milano, Italy.

Astrocytes are ubiquitous in the brain and have multiple functions. It is
becoming increasingly clear that they play an important role in monitoring the
neuromicroenvironment in CNS and in information processing or signaling in the
nervous system in normal conditions and respond to CNS injuries in a gradual
and varied way. It is still debated whether such reactions are beneficial or
detrimental. It was believed that reactive astrogliosis observed in most
neurological disorders may regulate the removal of toxic compounds produced by
damaged neurons and support neuronal growth by releasing trophic factors.
However it was also suggested that astrocytes contribute to a decline of
neurologic function, for example by accumulation and release of excitotoxic
aminoacids after ischemia and oxidative stress, formation of epileptogenic
scars in response to CNS injury and metabolism of protoxins to potent toxins.
In a number of metabolic diseases astrocytes, not neurons, may be the primary
target. The astrocyte's role in normal and pathological conditions will be
discussed in the light of recent information about their metabolism, receptor
distribution and release.

8e. Neurotoxicology 1998 Feb;19(1):7-17; discussion 37-8 
Astrocytic functions and physiological reactions to injury: the potential to
induce and/or exacerbate neuronal dysfunction--a forum position paper.
Aschner M
Department of Physiology and Pharmacology, Bowman Gray School of Medicine, Wake
Forest University, Winston-Salem, NC 27157-1083, USA.

This forum highlights the wide diversity of astrocytic functions which maintain
CNS homeostasis, well beyond the originally proposed passive cytoskeletal
support role for these cells. Astrocytic potential in modulating damage and
repair is also reflected in this forum. While astrocytes may potentially play
a primary role in epilepsy, and in neurodegenerative disorders such as
Huntington's diseases, HIV, and demyelination, one needs to keep in mind that
to a large extent evidence supporting involvement of astrocytes in these
diseases is derived from in vitro studies. Observations on regional
heterogeneity and functional specialization of astrocytes also suggest that
astrocytes have adapted to perform functions specific to their respective
residence site. Therefore, it is necessary to identify potentially damaging
consequences of astrocytic functions in vivo, although these analyses will be
undoubtedly extremely complex. Expanded investigations on astrocytic
involvement in neurotoxicity and neurodegeneration is clearly warranted, and as
new experimental tools are developed it is likely that further strides will be
made in our understanding of astrocyte functions, both in health and disease.

8f. Keio J Med 1997 Jun;46(2):55-60 
Neurotrophic activity in cytokine-activated astrocytes.
Yoshida K, Toya S
Department of Neurosurgery, School of Medicine, Keio University, Tokyo, Japan.

Accumulating evidence indicates that various neurotrophic factors (NTFs) exist
and function in the brain. In the mature mammalian brain, NTF expression is
exclusively restricted to neurons. However, astrocytes activated by various
cytokines, including fibroblast growth factor and interleukin-1 beta, produce
a significant amount of nerve growth factor (NGF) in vitro. Furthermore, non-NGF
type NTF expression in astrocytes is also activated by the cytokines. The
cytokines also enhance both release of ciliary neurotrophic factor from and
expression of high-molecular weight basic fibroblast growth factor (FGF) in
astrocytes. In the early phase following brain injury, cytokine-activated
astrocytes rescue the damaged neurons via NTFs and other biologically active
molecules.

9a. Int Immunol 1993 Nov;5(11):1445-50 
Polyclonal activation of immature B cells by preactivated T cells: the role of
IL-4 and CD40 ligand.
Brines RD, Klaus GG
Laboratory of Cellular Immunology, National Institute for Medical Research,
Mill Hill, London, UK.

It is well-established that preactivated CD4+ T cells can activate mature B
cells in a polyclonal, MHC-unrestricted fashion. We have used this system to
investigate the effects of T cell-derived signals on immature B cells purified
from the spleens of neonatal mice, since these cells are unresponsive to many
polyclonal activators and are exquisitely sensitive to tolerization. We show
that immature B cells can be induced to proliferate by anti-CD3 activated,
fixed Th1 and Th2 cells, although the latter induce a greater response than the
former. Antibodies to IL-4 partially blocked stimulation by Th2 cells, whereas
antibodies to IL-2 and IL-5 had no effect on responses to Th1 cells. This
suggested that molecules in addition to IL-4 contribute to the capacity of T
cells to induce B cell activation, one likely candidate being the ligand for
CD40. We therefore generated mouse erythroleukemia (MEL) transfectants which
express CD40 ligand (CD40L). These transfectants also induced proliferation of
immature B cells, which is enhanced by IL-4. Unlike the situation with mature
B cells, both anti-mu and anti-delta antibodies inhibited the activation of
immature B cells by CD40L-MEL cells. However, this inhibition was reversed by
IL-4, which synergized with signals delivered through CD40 to render immature
B cells refractory to negative signals delivered through sIg. Taken together
these data suggest that immature B cells can be activated by T cell-derived
contact signals and that CD40L-CD40 interactions, in the presence of IL-4, are
capable of abrogating the negative signals generated via sIgM and sIgD
receptors expressed by these cells.
PMID: 7505108, UI: 94083372


9b. Clin Immunol Immunopathol 1989 Sep;52(3):386-91 
Interleukin 2 as an inducer of proliferation in Klebsiella-stimulated human B
cells.
Stuber F, Ernst M, Flad HD, Gross WL

I. Medizinische Universitatsklinik, Kiel, Federal Republic of Germany.
Klebsiella membranes (KLEBS M), a membrane preparation from Klebsiella
pneumoniae, acts as a T-cell independent polyclonal B-cell activator (PBA),
stimulating B cells to mature into Ig-secreting cells without prior
proliferation. The KLEBS M-activated B cells express Tac antigen. Recombinant
interleukin 2, used in costimulation with KLEBS M, induces proliferation and
increases the Ig secretion of B cells. On the other hand, KLEBS M-activated B
cells do not respond to human BCGF. The lack of proliferation in KLEBS
M-stimulated mononuclear cells is shown to be due to the inability of this PBA
to induce the production of interleukin 2 in T cells.
PMID: 2667821, UI: 89337209


9c. Dtsch Med Wochenschr 1989 Jul 14;114(28-29):1101-6 
[Polyclonal B-cell activation by so-called immunopotentiation].
[Article in German]
Kekow J, Hobusch G, Ludemann J, Gross WL
I. Medizinische Universitatsklinik Kiel.

Immuno-augmentation with substances of bacterial origin was studied in vitro
for its ability to induce polyclonal B-cell activation. Biostim, Broncho-Vaxom,
Omnadin, Paspat and OK 432 were compared for their B-cell mitogenicity with
classical polyclonal B-cell activators (Staph. aureus Cowan I, KlebsM, Pokeweed
mitogen). B-cell mitogenicity, as measured by 3H-thymidine incorporation into
proliferating blood B-cells, was not induced by any of the studied
preparations. On the other hand, OK 432 produced a T-cell dependent and Biostim
a T-cell independent blood B-cell differentiation in immunoglobulin producing
cells. However, the extent of immunoglobulin production was clearly less than
with the polyclonal B-cell activator KlebsM. These results demonstrate that, in
some of the preparations, in vivo polyclonal B-cell activation can be expected
to occur.
PMID: 2787237, UI: 89305238


9d. Int J Immunopharmacol 1988;10(4):347-52 
Immunoglobulin secretion of mononuclear cells induced by various mitogens.
Shoenfeld Y, Teplizki H, Buskila D, Luedemann J, Gross W
Department of Medicine, Corob Research Center, Soroka University Hospital,
Beer-Sheva, Israel.

Immunosufficiency can be evaluated by Ig secretion subsequent to mitogenic
stimulation of human mononuclear cells (MNC). It seems that there are
significant differences in immunoglobulin class secreted by these cells when
stimulated with various polyclonal activators. The aim of the current study was
to analyse these differences. MNC cells was randomly obtained from nine healthy
blood donors and were activated by Epstein-Barr virus (EBV), group-A
streptococcus (A-ScM), Staphylococcus aureus (SAC), Klebsiella pneumonia
(Kleb-M) and pokeweed mitogen (PWM). Significantly increased levels of IgM were
recorded after a 7 day incubation followed by stimulation with Kleb-M (6.2 +/-
2.9) and EBV (5.9 +/- 4.5) compared to inactivated MNC (1.6 +/- 1.4), and
following 10 days incubation then stimulation by EBV (13.4 +/- 5.5) and Kleb-M
(9.9 +/- 4.2) compared to unstimulated cells (2.9 +/- 1.8). Significantly
greater IgG levels were achieved following incubation with EBV (3.0 +/- 4.0)
and PWM (2.4 +/- 1.3) after 7 days (vs 0.6 +/- 0.4 in unstimulated cells) and
by PWM (11.7 +/- 5.3) and Kleb-M (8.8 +/- 3.9, vs 2.3 +/- 2.2) after 10 days.
The present data emphasize the significance of merging both mitogen selection
and culture duration for acquiring information and high fidelity results of
immunoglobulin secretion by polyclonal activators.
PMID: 2844683, UI: 89007253


9e. Clin Exp Immunol 1987 Oct;70(1):201-8 
Polyclonal activation of murine B cells by a membrane proteoglycan of
Klebsiella pneumoniae.
Millet I, Lafont S, de Fraissinette A, Jeannin M, Revillard JP, Normier G,
Dussourd d'Hinterland L
INSERM U.80, CNRS ERA 1177, Lyon, France.

The lymphocyte activating properties of a membrane proteoglycan (MPG) extracted
from a mutant non-encapsulated strain of Klebsiella pneumoniae (Kp) (biotype a
I-145) were investigated. Kp MPG induced a strong proliferative response of
BALB/c spleen cells and Peyer's patches cells. Thymidine incorporation was
dose-related (from 1 to 100 micrograms Kp MPG/ml) and reached a maximum at day
3. It was not reduced by removal of most adherent cells, nor by depletion of
Thy1-2 positive cells, but it was abrogated by removal of surface
immunoglobulin bearing cells. Spleen cells from nude mice and those from
C3H/Hej mice were strongly stimulated by Kp MPG. Conversely Kp MPG did not
induce interleukin 2 production and did not trigger the proliferation of
thymocytes but stimulated interleukin 1 production by adherent spleen cells.
Finally, unfractionated or B-enriched spleen cells cultured with Kp MPG
synthesized IgM and, to a lesser extent, IgG and IgA. It is concluded that Kp
MPG is a T-independent polyclonal B cell activator and an inducer of
interleukin 1 production.
PMID: 3319300, UI: 88081073


9f. Immunobiology 1985 Nov;170(4):293-304 
Cell wall preparation consisting of group A carbohydrate and peptidoglycan
moieties from Streptococcus pyogenes activates murine B lymphocytes.
Morisaki I, Kimura S, Torii M, Michalek SM, McGhee JR, Okahashi N, Hamada S

Cell walls from Streptococcus pyogenes strain Sv (Group A, M type 3) were lysed
with M1 endo-N-acetylmuramidase, and the group A-specific carbohydrate antigen
was purified by Sephadex G-100 gel filtration. The initial eluting antigen
(M1gA) peak was assessed for mitogenic and polyclonal lymphocyte-activating
properties in murine spleen cell cultures. Good mitogenic responses were
induced over a broad dose range (1-100 micrograms) with M1gA in both BALB/c and
C3H/HeJ splenic cultures. Similar mitogenic responses were induced in nude
(nu/nu) and nu/+ splenic cultures, suggesting that M1gA is a B cell mitogen.
The M1gA induced anti-trinitrophenyl, anti-sheep erythrocytes, and anti-horse
erythrocytes polyclonal plaque-forming cell responses in splenic cell cultures.
Studies with purified splenic B cells and M1gA suggest that the mitogenic
responses were indeed thymic independent. These studies clearly indicate that
native group A carbohydrate antigen is a B cell mitogen and polyclonal B cell
activator.
PMID: 3910560, UI: 86110332


9g. Cell Immunol 1985 Mar;91(1):52-9 
Potent adjuvant action of lipopolysaccharides possessing the O-specific
polysaccharide moieties consisting of mannans in antibody response against
protein antigen.
Kido N, Ohta M, Ito H, Naito S, Nagase F, Nakashima I, Kato N

It was previously reported that Klebsiella O3 lipopolysaccharide (LPS) exhibits
extraordinarily strong adjuvant activity in augmenting antibody response
against protein antigens in mice compared with other kinds of LPS, for example,
LPS from Escherichia coli O55, O111, and O127 and Salmonella enteritidis. The
present study was undertaken to clarify the relationship between the strong
adjuvant activity in augmenting antibody response against deaggregated bovine
gammaglobulin and the chemical structure of LPS. Among LPS from Klebsiella O1,
O4, O5, and O7, only O5 LPS exhibited nearly the same degree of the strong
adjuvant activity as did O3 LPS. The adjuvant activity of the other LPS was
very weak in a degree similar to that of LPS from E. coli O55 and O127. Even
when the natural forms of Klebsiella O3 LPS and O1 LPS were converted to
various defined uniform salt forms, their adjuvant activity did not
significantly differ from that of the respective natural forms. It is therefore
unlikely that the difference in strength of the adjuvant activity between
Klebsiella O3 LPS and O1 LPS is due to the difference in their salt forms. The
common feature in the structures of Klebsiella O3 LPS and O5 LPS is their
O-specific polysaccharide chains consisting of the mannose homopolysaccharides
(mannans). LPS from E. coli O8 and O9, the O-specific polysaccharide chains of
which consist of the mannans, also exhibited much stronger adjuvant activity
than do LPS from E. coli O55 and O127, and the strength of the adjuvant
activities of the former two was comparable to that of LPS from Klebsiella O3
and O5. On the other hand, LPS from Klebsiella O3 and O5 and E. coli O8 and O9
showed the ability to activate B lymphocytes polyclonally in vivo in a degree
similar to that of the other kinds of LPS. From the present results it can be
concluded that LPS possessing the O-specific polysaccharide moieties consisting
of the mannans exhibit extraordinarily strong adjuvant activity in augmenting
antibody response against protein antigen.
PMID: 2578897, UI: 85124650

9h. Immunol Lett 1985;10(3-4):213-6 
T-cell influence on the B-cell differentiation process induced by Klebsiella.
Gross WL, Sieg I, Steppat D

Klebsiella pneumoniae K43 cell membrane preparations (Klebs M) have been
characterized previously as a human polyclonal B cell activator (PBA) that
stimulates purified B cells to differentiate into immunoglobulin (Ig) secreting
cells with negligible prior or parallel proliferation and in the absence of T
cells. The aim of the present study was to define the cellular interactions in
the regulation of Klebs M induced B-cell differentiation. For this purpose
OKT4+ and OKT8+ cell populations were negatively selected with reasonable
purity by means of a panning technique or by complement-mediated cytolysis
using monoclonal OKT4 and OKT8 antibodies. The resulting cell populations were
added to purified autologous B cells exposed to Klebs M or, as a control,
pokeweed mitogen (PWM). In the Klebs M system both the OKT4+ and the OKT8+ cell
subsets markedly enhanced IgM production; however, the helper effect of the
OKT4+ cell subset was much more intense than that of the OKT8+ subset. In the
PWM system only the OKT4+ cells provided help for B-cell differentiation. The
OKT8+ subset demonstrated suppressor activity in the presence of an adequate
helper cell (OKT4+ subset) function. These results indicate that Klebs M
behaves like a "relatively T cell-independent PBA".
PMID: 2931358, UI: 86006921


9i. J Immunol 1984 Feb;132(2):616-21 
Influence of RU 41.740, a glycoprotein extract from Klebsiella pneumoniae, on
the murine immune system. I. T-independent polyclonal B cell activation.
Wood CD, Moller G

A glycoprotein extract from Klebsiella pneumoniae, RU 41.740, has been shown
clinically to reduce infectious episodes in patients prone to frequent
infections, and experimentally to reduce mortality in animals infected with
various bacteria or viruses. The method of action of RU 41.740 in conferring
protection from infection is as yet unknown, but in experimental animals, RU
41.740 appears to influence B and T cells as well as macrophages. We report
here that RU 41.740 is a strong polyclonal B cell activator and that it
activates B cells in a T-independent manner. RU 41.740 also activates spleen
cells from the LPS-nonresponder mouse strains, C3H/HeJ and C57BL/10ScCr, which
indicates that the polyclonal B cell activating ability of RU 41.740 is not due
to contamination by LPS.
PMID: 6197468, UI: 84088971


9j. Clin Immunol Immunopathol 1983 May;27(2):261-71 
Polyclonal activation of immunoglobulin secretion without prior DNA synthesis
in human B lymphocytes induced by Klebsiella pneumoniae.
Gross WL, Rucks A, Hahn G, Ullmann U

The capacity of various cell preparations of Klebsiella pneumoniae K43 (Klebs)
to induce [3H]thymidine uptake and immunoglobulin (Ig) secretion by human
mononuclear blood cells (MNC) and their lymphocyte subpopulations was
investigated. All Klebs preparations were virtually devoid of mitogenic
properties, in contrast to control preparations of pokeweed mitogen (PWM) and
group A streptococcal cell membranes (A-ScM). Klebs induced differentiation of
B cells into Ig-secreting cells. B-cell populations that were sufficiently
depleted of T cells to be unresponsive to A-ScM ("highly purified B cells")
showed a marked response to Klebs. Similarly, the number of plaque-forming
cells (PFC) in Klebs-driven cultures did not change after restitution of T
cells, whereas the presence of restituted T cells augmented the B-cell response
to PWM and A-ScM. Radical removal of adherent MNC ("monocytes"), however,
completely abrogated the PFC response and [3H]thymidine uptake of both MNC
activated by Klebs and MNC activated by PWM or A-ScM.
PMID: 6191901, UI: 83260061


9k. Clin Exp Immunol 1983 May;52(2):372-80 
Klebsiella pneumoniae stimulate highly purified human blood B cells to mature
into plaque forming cells without prior proliferation.
Gross WL, Rucks A

The plaque forming cell (PFC) and proliferative responses of human peripheral
blood lymphocytes and highly purified blood B cells induced by pokeweed mitogen
(PWM) and group A streptoccocal cell membranes (A-ScM) were compared with the
responses triggered by various cell preparations of Klebsiella pneumoniae K 43
(Klebs). The number of PFC was determined by a protein A plaque assay, and
lymphoproliferation was measured by 3H-thymidine incorporation. In cell
cultures stimulated with PWM and A-ScM, lymphocyte proliferation appeared to be
associated with the generation of PFC. Klebs caused development of PFC without
measurable prior proliferation. Whereas the response to PWM and A-ScM was
absolutely T cell-dependent, highly purified B cells generated PFC when
incubated with Klebs. Moreover, restitution of T cells to the B cell fraction
did not augment (or diminish) the number of plaques. These studies establish
that Klebs cell envelope structures contain a T cell-independent polyclonal B
cell activator for human B lymphocytes in a high stage of differentiation. Use
of this probe should provide further insight into the cellular interactions
involved in the differentiation of antibody forming cells in humans.
PMID: 6190601, UI: 83233332


9l. Microbiol Immunol 1980;24(2):141-54 
Adjuvant action of capsular polysaccharide of Klebsiella pneumoniae on antibody
response. VIII. Its effect on the size and the number of cells of regional
lymph node and other lymphoid organs.
Yokochi T, Nakashima I, Kato N

Changes in the number of cells and the weight of various lymphoid organs of
mice, such as the regional lymph node (right inguinal node), spleen, thymus,
bone marrow, and peripheral blood, were followed after the subcutaneous
injection of the capsular polysaccharide of Klebsiella pneumoniae (CPS-K). For
comparison, the changes after injection of various polyclonal lymphocyte
activators (PLA) including various preparations of bacterial lipopolysaccharide
(LPS) were concurrently studied. The number of cells of all of the lymphoid
organs tested and that of nucleated cells in the peripheral blood decreased
significantly within a few days after injection of CPS-K, and increased later.
Above all, the increase in the number of cells and in the weight of the
regional lymph node was most prominent (about 10 times larger than that of the
normal control). Such a marked increase in the number of cells of the regional
lymph node was not induced by the injection of any preparation of LPS or any
other PLA tested. The initial decrease in the number of cells after CPS-K
injection was most marked and long lasting in the thymus. Although LPS prepared
by Westphal's method from Escherichia coli 055 or Salmonella enteritidis
exhibited a stronger decreasing effect on the number of cells of the thymus,
the effect of LPS prepared by Westphal's method from E. coli O111 or that by
Boivin's method from E. coli O55 was similar to that of CPS-K. It is concluded
therefore that CPS-K has the ability to decrease the number of cells of various
lymphoid organs, especially that of the thymus, initially after injection,
which is a property in common with LPS, and CPS-K has a unique ability to
increase markedly the cells of various lymphoid organs, especially those of the
regional lymph node, at later stages after injection. Considering that CPS-K
exhibits a much stronger adjuvant effect on the antibody response than does LPS
or other polyclonal lymphocyte activators, it is suggested that this
extraordinarily potent activity of CPS-K in increasing the number of cells of
the regional lymph node is closely related to its strong adjuvant action.
PMID: 6991886, UI: 80209292

10a. Infect Immun 1993 Mar;61(3):852-60 
C1q binding and activation of the complement classical pathway by Klebsiella
pneumoniae outer membrane proteins.
Alberti S, Marques G, Camprubi S, Merino S, Tomas JM, Vivanco F, Benedi VJ
Universidad de las Islas Baleares, Departamento de Biologia Ambiental, Palma de
Mallorca, Spain.

The mechanisms of killing of Klebsiella pneumoniae serum-sensitive strains in
nonimmune serum by the complement classical pathway have been studied. The
bacterial cell surface components that bind C1q more efficiently were
identified as two major outer membrane proteins, presumably the porins of this
bacterial species. These two outer membrane proteins were isolated from a
representative serum-sensitive strain. We have demonstrated that in their
purified form, they bind C1q and activate the classical pathway in an
antibody-independent manner, with the subsequent consumption of C4 and
reduction of the serum total hemolytic activity. Activation of the classical
pathway has been observed in human nonimmune serum and agammaglobulinemic serum
(both depleted in factor D). Binding of C1q to other components of the
bacterial outer membrane, in particular the rough lipopolysaccharide, could not
be demonstrated. Activation of the classical pathway by this lipopolysaccharide
was also much less efficient than activation by the two outer membrane
proteins. The antibody-independent binding of C1q to serum-sensitive strains
was independent of the presence of capsular polysaccharide, while strains
possessing lipopolysaccharide O antigen bind less C1q and are resistant to
complement-mediated killing.
PMID: 8432605, UI: 93162823

10b. Mol Immunol 1994 Nov;31(16):1239-46 
Acylation of the lipid A region of a Klebsiella pneumoniae LPS controls the
alternative pathway activation of human complement.
Mey A, Ponard D, Colomb M, Normier G, Binz H, Revillard JP
Laboratoire d'Immunologie INSERM U80 Lyon, France.

Two mechanisms of direct activation of the complement system by LPS have been
extensively documented: (i) activation of the alternative pathway (AP) by the
polysaccharide region, and (ii) activation of the classical pathway (CP) by the
lipid A region. Here we demonstrate that LPS from the Klebsiella pneumoniae
I-145 strain activates the AP by a mechanism dependent on the acylation of the
lipid A region. Cleavage of C3 by K. pneumoniae LPS in EGTA was blocked by
polymyxin B. Two 34 kDa derivatives were prepared from a membrane extract of
this K. pneumoniae strain: (i) an acyl-poly (1,3) galactoside containing two
galactosamine-bound ester-linked and two amide-linked fatty acids (EFA-APG),
and (ii) an acyl-poly (1,3) galactoside devoid of ester-linked fatty acids
(APG). APG and EFA-APG share the structure of LPS molecules, with a long
polysaccharidic chain, a core, and a lipid A region. The AP was activated by
EFA-APG but not by APG nor by the isolated polygalactose chain GC-APG,
indicating a critical role for ester-linked fatty acids in AP activation.
Polymyxin B which binds to the lipid A region of LPS completely inhibited AP
activation by EFA-APG. A small part of EFA-APG was shown to form aggregates in
saline, but aggregation was not decreased by polymyxin B. Furthermore, APG
formed aggregates of similar size although it was not able to activate AP.
Therefore the role of lipid A acylation in triggering AP activation is not
exclusively mediated by aggregation of the molecule. LPS from the rough strain
of Salmonella minnesota (Sm Re LPS) directly activates the CP but not the AP.
However, when mixed with the polygalactose chain GC-APG, Sm Re LPS activated
the AP. The data demonstrate a cooperation between the lipid A region and the
polysaccharidic chain in activation of the AP. Similar cooperation may occur
with other LPS molecules.
PMID: 7969185, UI: 95059081


10c. Chemotherapy 1997 Mar-Apr;43(2):132-6 
Binding of complement C3 to Klebsiella pneumoniae treated with sub-MIC
cefodizime and chemiluminescence response of human polymorphonuclear
leukocytes.
Nomura S, Kuroiwa A, Murata K, Nagayama A
Department of Microbiology, Fukuoka University School of Medicine, Japan.

The binding of complement C3 to the cell surface of Klebsiella pneumoniae
exposed to human serum complement after treatment with or without sub-MIC of
antibiotics was examined by double diffusion immunoprecipitation against
anti-human complement C3, and the production of oxygen-derived radicals by
human polymorphonuclear leukocytes stimulated by complement-opsonized K.
pneumoniae after treatment with or without sub-MIC of antibiotics was measured
using the chemiluminescence (CL) assay. Complement C3 bound to the cell surface
of K. pneumoniae treated with cefodizime was detected after exposure to human
serum complement. The CL response induced by complement-opsonized bacteria
after treatment with cefodizime was much higher than the response induced by
nontreated bacteria or complement-opsonized bacteria after treatment with other
antibiotics. These findings indicate that treatment with sub-MIC cefodizime
make K. pneumoniae more susceptible to opsonization by complement and promotes
the specific phagocytosis mediated by complement receptors.
PMID: 9084922, UI: 97239260

10d. J Immunol 1990 Apr 15;144(8):3106-10 
Strong interaction of lipopolysaccharides possessing the mannose
homopolysaccharides with complement and its relation to adjuvant action.
Yokochi T, Inoue Y, Kimura Y, Kato N
Department of Microbiology, Fukui Medical School, Japan.

LPS from Klebsiella pneumoniae O3 (KO3 LPS) exhibited an extremely high
anticomplementary activity by the hemolysis assay using human sera. The free
lipid A isolated from KO3 LPS by acid hydrolysis and R form LPS from a mutant
lacking the O-specific polysaccharide portion possessed lower anticomplementary
activity, and the O-specific polysaccharide fraction isolated from KO3 LPS
alone did not activate the C system. It was suggested that the O-specific
polysaccharide moiety enhanced the C activation by the lipid A portion. This
was also supported by the finding that modification of the O-specific
polysaccharide moiety with Con A or tyramine decreased anticomplementary
activity of KO3 LPS, and that the other LPS preparations possessing the mannose
homopolysaccharides as the O-specific polysaccharide portions such as KO3 LPS,
such as LPS from Klebsiella O5, Escherichia coli O8 and O9, exhibited a high
anticomplementary activity. KO3 LPS could activate the C system in either the
classical or the alternative pathway, whereas the lipid A or R form LPS
activated the classical pathway alone. The intensity of anticomplementary
activity of LPS was parallel to that of their adjuvant action on antibody
response to deaggregated BSA. The role of the anticomplementary activity in the
expression of the adjuvant action of LPS is discussed.
PMID: 2182713, UI: 90217520