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]  Kp is associated with distended abdomen (eb, 1a-1b); although other causes
known.
[b]  Intraperitoneal LPS can reduce CD5+ cells; and Kp LPS affects
intraperitoneal macrophages by a calcium-dependent process (2a; 2b).
[c]  Peritoneal tissues may have reduced activation of complement:
          "The results obtained could be regarded as a consequence of
          the lowered alternative pathway serum complement activity
          and the crucial role of the diminished level of complement
          in the peritoneal cavity." (3).
[d]  Lipoproteins can alter inflammatory responses to Kp and other bacteria
without altering the underlying infection (4a); and fish oils have been shown
to be helpful (4b).
[e]  Vitamin B2 enhances immunity against Kp (5), and other drugs may also be
helpful (eg, cefodizime; 6a); and cedofizime has been particularly effective
within the peritoneum (6b), as have certain third-generation cephalosporins
(6c); other substances may now be available for drug-resistant Kp species (eg,
6d); and combination treatments can be more effective (eg, 6e-f).

[f]  Other immune modulators can be effective (7a-d).
[g]  Histamine can be released by Kp (8).
[h]  Cytokines (eg, IL-8) can be helpful or harmful, depending upon the immune
status of the individual (9a); type-I interferon can be helpful against Kp (9b),
as can recombinant IL-2 (9c).
[i]  Kp has iron-regulated outer-membrane proteins; and has strains described
as virulent or avirulent (though I do not know the extent to which this
distinction occurs in wild type strains); and Kp may have intrinsic means to
evade complement-mediated killing (10).
[j]  Iron depletion or excess alters Kp responses in the presence of
cephalosporins (11a), noting that Michael's WBC/RBC evaluations show atypical
iron/hemoglobin values; and altered iron may affect and be a sign of E. coli
colonization (11b). 

[k]  Certain molecules from Kp enhance immunological killing of Candida (12),
and this would be consistent with Michael's seldom noted intestinal Candida
while having a high level of Kp colonization. 
[l]  CMV exacerbates Kp (13), which is at least theoretically important to
Michael because neonatal CMV can be asymptomatic, can affect the
gastrointestinal tract, impairs antibody responses against CMV (14a-b), and can
be present in individuals having no detectable anti-CMV antibodies (14c), which
justifies using PCR of peripheral monocytes so as to rule out CMV as an
etiologically significant factor in Michael, especially since CMV has been shown
to impair host immune responses against Kp (14d).
[m]  Kp impairs immunity against other bacterial infections (15a-c).
[n]  If systemic Kp and/or hEc fragments are reaching cells of the blood-brain
barrier, then vascular inflammation could be occurring (16a-b), along with
associated hypoperfusion and hypofuction in adjoining brain regions and neuronal
pathways connected thereto (17), as further delineated on a webpage
http://www.jorsm.com/~binstock/bbb-inf.htm

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

1. Ryumachi 1998 Dec;38(6):825-30 
[Pneumoperitoneum with systemic sclerosis].
[Article in Japanese]
Nakajima A, Koseki Y, Suwa A, Imaeda H, Okamura T, Tsutsumino M, Sendo W, Terai
C, Hara M, Kashiwazaki S, Inada S
Division of Rheumatic Diseases, Tokyo Metropolitan Ohtsuka Hospital.

We report a case of systemic sclerosis (SSc) complicated with benign
pneumoperitoneum without apparent pneumatosis cystoides intestinalis (PCI). A
43-year-old woman was admitted to our hospital because of prominent abdominal
distension in April 1997. Raynaud's phenomenon has been detected since 1991.
She was suffering from recurrent diarrhea, constipation, and subileus. The
diagnosis of SSc was made in 1996 based on the sclerosis in her face, forearms,
and chest, and hypomotility of the esophagus. On admission, she presented no
signs of peritoneal irritation. The laboratory data revealed that white blood
cell count was 7,400/mm3 and C-reactive protein was 0.1 mg/dl. Chest and
abdominal roentgenograms showed massive free air under the diaphragm,
dilatation of small and large intestine, and air-fluid level. PCI was not
apparent. Pneumoperitoneum was improved after four weeks with intravenous
hyperalimentation. But she presented recurrent severe diarrhea and high fever
whenever she tried to take food orally. Klebsiella pneumoniae was proved in her
jejunal juice by bacteriologic examination. Intravenous prostaglandin F2 alpha
and oral fosfomycin calcium intake made her condition better. Benign
pneumoperitoneum without PCI is rarely reported in the patients with SSc. In
her condition, weakness of intestinal wall, hypomotility of intestine, unusual
bacterial overgrowth, and elevated intraluminal pressure made intraluminal gas
go through the wall of the fragile intestine of SSc. As operation of intestine
of SSc usually cause miserable outcome, pneumoperitoneum accompanied with SSc
even if PCI is apparent or not must be treated with conventional manner while
there is no signs of peritoneal irritation.
Publication Types:  Review  Review of reported cases
PMID: 10047721, UI: 99157251

1b. Indian J Pathol Microbiol 1998 Jan;41(1):101-2 
Intraperitoneal abscess in a new born secondary to Klebsiella septicaemia--a
case report.
Kumari PG, Rao PL, Antony B, Shivananda PG
Department of Microbiology, Kasturba Medical College, Manipal.

A five day old female baby was admitted with distension of abdomen since birth
and nonbilious vomiting, fever of one day duration. Blood culture grew
Klebsiella pneumoniae. Abdominal exploration revealed thick walled cavity
containing purulent fluid grew klebsiella pneumoniae which was sensitive to
various antibiotics including gentamycin. The child was treated with injection
gentamycin and ceftazidime. The child had uneventful recovery and is doing well
3 years post operatively.
PMID: 9581084, UI: 98242185


2a. Infect Immun 1997 Jan;65(1):122-6 
Marked reduction of mouse peritoneal CD5+ B cells by intraperitoneal
administration of lipopolysaccharide.
Paeng N, Kido N, Kato Y, Sugiyama T, Koide N, Naruse M, Jiang GZ, Lwin T,
Yoshida T, Yokochi T
Department of Microbiology and Immunology, Aichi Medical University, Nagakute,
Japan.

Intraperitoneal administration of lipopolysaccharide to mice induced a marked
reduction of CD5+ B cells in the peritoneal cavity. The reduction was not
induced by intravenous, subcutaneous, or oral administration of
lipopolysaccharide. The reduction continued for about 10 days after the
injection, and the CD5+ B-cell count recovered to the normal state about 14
days after the injection. The reduction of peritoneal CD5+ B cells might be
caused by apoptotic cell death. Injection of lipopolysaccharide did not result
in production of antibody to lipopolysaccharide. On the other hand,
intraperitoneal injection of heat-killed bacteria did not induce a reduction of
peritoneal CD5+ B cells and elicited the definite production of antibody to
lipopolysaccharide.
PMID: 8975901, UI: 97130025

2b. Vopr Med Khim 1993 Mar-Apr;39(2):58-62 
[Induction of Ca2+-dependent chemiluminescent response of peritoneal
macrophages with bacterial lipopolysaccharides].
[Article in Russian]
Uvarov VD, Shepelev AP, Poliakov VM

A lipopolysaccharide preparation obtained from Klebsiella pneumonia was shown
to affect primarily the peritoneal macrophages after intraperitoneal
administration. Transition of the macrophages to a new metabolic state in
response to opsonized zymosan was responsible for a distinct increase in the
rate of luminol-dependent chemoluminescence. At the same time, the macrophages
produced chemiluminescence was only slightly increased in the presence of Ca2+
ionophore A23187. These dissimilar alterations observed suggest that the
primary reaction of macrophages in response to the lipopolysaccharide occurred
with the increase in content of intracellular free calcium. Administration of
finoptin (calcium influx inhibitor) into suspension of macrophages caused a
decrease in the rate of chemiluminescence response.
PMID: 8511894, UI: 93289738


3. Int J Immunopharmacol 1996 Aug-Sep;18(8-9):515-9 
Correlation between inhibited alternative complement activity and the
protective effect induced by Nocardia lysozyme digest (NLD) during Klebsiella
pneumoniae infection in mice.
Ivanovska N, Georgieva P, Barot-Ciorbaru R
Department of Immunology, Bulgarian Academy of Sciences, Sofia.

Nocardia lysozyme digest (NLD) was administered intraperitoneally (i.p.) at a
dose of 500 micrograms/kg to normal and immunosuppressed mice for 3 consecutive
days prior to inoculation with Klebsiella pneumoniae. A protective effect was
observed when the pathogen was injected subcutaneously and intravenously, as
opposed to an aggravating effect obtained in the case of intraperitoneal
inoculation The i.p. administration of NLD partially restored the
immunosuppression caused by cyclophosphamide but did not change cobra
venom-induced deterioration of the infection. The results obtained could be
regarded as a consequence of the lowered alternative pathway serum complement
activity and the crucial role of the diminished level of complement in the
peritoneal cavity.
PMID: 9023591, UI: 97176017


4a. J Clin Invest 1996 Mar 15;97(6):1366-72 
Low-density lipoprotein receptor-deficient mice are protected against lethal
endotoxemia and severe gram-negative infections.
Netea MG, Demacker PN, Kullberg BJ, Boerman OC, Verschueren I, Stalenhoef AF,
van der Meer JW
Department of Medicine, University Hospital Nijmegen, The Netherlands.

Lipoproteins can bind lipopolysaccharide (LPS) and decrease the LPS-stimulated
production of pro-inflammatory cytokines. We investigated the effect of
increased plasma concentrations of low-density-lipoproteins (LDL) on survival
and cytokine production after a lethal challenge with either LPS or live
Gram-negative bacteria in LDL receptor deficient mice (LDLR-/-). The LDLR-/-
mice challenged with LPS had an eightfold increased LD50 when compared with the
wild type controls (C57Bl/6J), while tumor necrosis factor alpha (TNFalpha) and
interleukin-1 alpha (IL-1 alpha) plasma concentrations were decreased twofold.
LDLR-/- mice had significantly lower and delayed mortality than control mice
after infection with Klebsiella pneumoniae. 
     No differences in the outgrowth of bacteria in the organs were
     present between the two groups, while circulating cytokine
     concentrations were decreased twofold in LDLR-/- mice. 
In contrast, the LPS-stimulated in vitro production of cytokines by peritoneal
macrophages of LDLR-/- mice was significantly increased compared with controls.
This increase was associated with enhanced specific binding of LPS to the
macrophages of LDLR-/- mice. In conclusion, endogenous LDL can protect against
the lethal effects of endotoxin and Gram-negative infection. At least part of
this protection is achieved through decreased in vivo production of
pro-inflammatory cytokines, in spite of increased cytokine production capacity.
PMID: 8617867, UI: 96195732


4b. J Infect Dis 1992 May;165(5):898-903 
Dietary fish-oil supplementation in experimental gram-negative infection and in
cerebral malaria in mice.
Blok WL, Vogels MT, Curfs JH, Eling WM, Buurman WA, van der Meer JW
Department of General Internal Medicine, University Hospital Nijmegen,
Netherlands.

Dietary fish-oil supplementation interferes with eicosanoid production and
appears to decrease production of interleukin-1 (IL-1) and tumor necrosis
factor (TNF). The effect of fish oil was investigated in an intramuscular
Klebsiella pneumoniae infection in Swiss mice and in cerebral malaria induced
by Plasmodium berghei in C57B1/6 mice. After a low inoculum of K. pneumoniae,
90% of fish oil-fed mice survived; survival in control mice fed equal amounts
of corn or palm oil or normal chow was 30%, 40%, and 0, respectively. Cerebral
malaria occurred in only 23% of fish oil-fed mice; in the controls, cerebral
malaria developed in 61%, 81%, and 78%, respectively. Contrary to what was
expected, lipopolysaccharide-induced ex vivo production of IL-1 alpha and TNF
alpha by peritoneal cells was significantly enhanced in fish oil-fed mice
compared with controls. Indomethacin treatment did not alter the outcome in
these two infections, thus arguing against reduced prostaglandin synthesis as
an explanation for the increase in resistance to infection.
PMID: 1569340, UI: 92235493



5. J Vet Med Sci 1995 Aug;57(4):599-602 
Enhancement of resistance to bacterial infection in mice by vitamin B2.
Araki S, Suzuki M, Fujimoto M, Kimura M
Research and Development Division, Eisai Co., Ltd., Ibaraki, Japan.

We found that the intramuscular injection of vitamin B2 enhanced host
resistance to E. coli infection in a dose-dependent manner (6.25 mg/kg-100
mg/kg). Furthermore, VB2 exhibited the protective activity against Pseudomonas
aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, and Actinobacillus
pleuropneumoniae. The mechanism of action of VB2 for enhancing resistance in
mice may be, at least in part, its ability to stimulate the multiplication of
neutrophils and monocytes, and to activate macrophages.
PMID: 8519884, UI: 96081286


6a. Chemotherapy 1995 Jul-Aug;41(4):267-75 
Mechanism of enhancement of bactericidal activity of phagocytes against
Klebsiella pneumoniae treated with subminimal inhibitory concentrations of
cefodizime.
Nomura S, Nagayama A
Department of Microbiology, School of Medicine, Fukuoka University, Japan.

The effects of a sub-MIC of cefodizime on the morphology of the capsular
structures and on the surface physicochemical properties, such as
hydrophobicity and charge, of encapsulated Klebsiella pneumoniae were studied.
The enhancement of bactericidal activity of macrophages against bacteria
treated with sub-MICs of antibiotics was evaluated as the killing index.
Cefodizime treatment gave the highest value of 32. Electron microscope
observations revealed that the capsular material layer of cefodizime-treated K.
pneumoniae was markedly thinner (32 nm) than that of untreated bacteria (160
nm) or bacteria treated with other antibiotics (75-90 nm). Contact angle
measurement revealed that the surface of cefodizime-treated K. pneumoniae was
more hydrophobic than that of untreated bacteria or bacteria treated with other
antibiotics. Furthermore, the negative charge of the surface of K. pneumoniae
decreased significantly with cefodizime treatment compared with the surface of
untreated bacteria. These findings suggest that the treatment of K. pneumoniae
with a sub-MIC of cefodizime reduced the thickness of the capsular material
layer and that these changes increased the surface hydrophobicity of the
bacteria and decreased the negative charge of the bacterial surface to render
K. pneumoniae more susceptible to phagocytic activity by reducing the physical
repulsion between the bacteria and phagocytes.
PMID: 7555207, UI: 96045139

6b. Chemotherapy 1995 May-Jun;41(3):178-86 
In vitro and in vivo enhancement of bactericidal activity of phagocytes against
Klebsiella pneumoniae treated with subminimal inhibitory concentrations of
cefodizime.
Nomura S, Nagayama A
Department of Microbiology, School of Medicine, Fukuoka University, Japan.

The effects of a subminimal inhibitory concentration (sub-MIC) of cefodizime on
the bactericidal activity of phagocytes against encapsulated Klebsiella
pneumoniae were studied in an in vitro system using an established mouse
macrophage cell line, and in an in vivo system using mice in which many
phagocytes were induced in the peritoneal cavity. In the in vitro system, the
bactericidal activity of mouse macrophages against K. pneumoniae treated with
a sub-MIC of cefodizime was significantly enhanced, and was greater than that
of cefotaxime or cefoperazone. Significantly more bacteria treated with a
sub-MIC of cefodizime were killed by serum complement than those treated with
cefotaxime or cefoperazone. In the in vivo system, cefodizime-treated bacteria
were phagocytosed and killed by phagocytes in the mouse peritoneal cavity,
whereas, cefotaxime- and cefoperazone-treated and untreated bacteria were hardly
phagocytosed at all or killed by phagocytes in the mouse peritoneal cavity, and
bacterial regrowth was observed 24 h after bacterial challenge. Furthermore, the
virulence of K. pneumoniae in mice was reduced more by treatment with cefodizime
than with cefotaxime or cefoperazone. These findings indicate that K. pneumoniae
treated with a sub-MIC of cefodizime become more susceptible to the bactericidal
activity of phagocytes both in vitro and in vivo. This provides evidence that
cefodizime at a sub-MIC may act together with the phagocytes against bacterial
infections.
PMID: 7656663, UI: 95385453

6c. Chemotherapy 1989;35(5):326-9 
Penetration of third-generation cephalosporins into human peritoneal tissue.
Berger SA, Dan M, Serour F, Gorea A, Levenberg A, Krispin M
Department of Microbiology, Tel-Aviv Medical Center, Israel.

Each of 40 patients underwent elective laparotomy following administration of
a single 1.0-gram intravenous dose of ceftizoxime, ceftriaxone, cefoperazone or
cefotaxime. Therapeutic concentrations of cefoperazone and ceftriaxone were
achieved in peritoneal tissue in 20/20 patients. Only 9/20 samples from
patients receiving the other two antibiotics had detectable antibiotic
activity. The antibiotic concentration in peritoneal fluid (7 samples) was
2.36-11.15 times higher than that of concurrently obtained peritoneal tissue.
When adjusted for the in vitro susceptibility (MIC) of potential peritoneal
pathogens, our data suggest that ceftriaxone and cefoperazone may be preferable
to other third-generation cephalosporins for the prophylaxis and therapy of
intraabdominal infection.
PMID: 2676403, UI: 90004579


6d. J Antibiot (Tokyo) 1986 Oct;39(10):1450-60 
The antibacterial activity of ticarcillin/clavulanic acid (BRL28500) against
ticarcillin-resistant bacteria.
Kasai T, Nishino T, Kazuno Y, Tanino T

The efficacy of BRL28500, a formulation of ticarcillin (TIPC, 15 parts) and
clavulanic acid (CVA, 1 part), against TIPC-resistant strains of Staphylococcus
aureus, Escherichia coli and Klebsiella pneumoniae was studied both in vitro
and in vivo. The MICs of BRL28500 against these beta-lactamase producing
strains were lower than those of TIPC or CVA alone against such strains. When
BRL28500 was added during the logarithmic growth phase of bacteria at a
concentration equivalent to the MIC, it demonstrated marked lytic activity.
Cells treated with BRL28500 underwent morphological change, becoming
filament-like, similar to those treated with TIPC alone. With CVA alone at
concentrations above the MIC the cells assumed a stable round form. In
bacterial cultures of the beta-lactamase-producing strains, TIPC was protected
from hydrolysis by the presence of CVA. The in vivo activity of BRL28500
against experimental infections in mice caused by beta-lactamase-producing
strains of bacteria was superior to that of TIPC alone. TIPC and CVA were found
to be well distributed in peritoneal fluid following subcutaneous
administration of BRL28500 into mice with peritoneal infections. The residual
TIPC concentrations achieved were higher than when TIPC alone was administered.
These results suggest that BRL28500 will be effective in the treatment of human
infections due to TIPC-resistant bacteria.
PMID: 3536827, UI: 87056659

6e. Am J Med 1986 Jun 30;80(6B):138-42 
Combination therapy: a way to limit emergence of resistance?
Michea-Hamzehpour M, Pechere JC, Marchou B, Auckenthaler R

The ability of antibiotic combinations to limit the emergence of resistance
during therapy was evaluated in a murine model. Peritonitis was produced by
injecting a mixture containing 10(8) colony-forming units of bacteria and
sterilized talcum into the peritoneum. Two hours later, a single antibiotic
dose was administered subcutaneously. The next day, peritoneal bacterial
populations were analyzed on Szybalski's gradients. Acquired resistance was
recorded when there was at least a fourfold increase in minimum inhibitory
concentrations compared with untreated animals. No resistance emerged after
amikacin monotherapy (15 mg/kg); however, resistance was frequently observed
after monotherapy with ceftriaxone (50 mg/kg) or pefloxacin (25 mg/kg).
Resistance to ceftriaxone and pefloxacin emerged, respectively, in 15 percent
and 83 percent of animals with Klebsiella pneumoniae, 71 percent and 54 percent
with Enterobacter cloacae, 0 percent and 83 percent with Serratia marcescens,
25 percent and 100 percent with Pseudomonas aeruginosa, and 0 percent with both
Escherichia coli and Staphylococcus aureus. 
     In mice with K. pneumoniae or E. cloacae infections, any dual
     combination of amikacin, pefloxacin, and ceftriaxone produced less
     acquired resistance than did monotherapy. 
In these animals, the combination of ceftriaxone and pefloxacin abolished all
resistance, whereas the combinations of amikacin plus ceftriaxone or amikacin
plus pefloxacin reduced the frequency of resistance by more than half. In
animals with P. aeruginosa or S. marcescens infections, resistance to
pefloxacin diminished or disappeared after treatment with the combinations of
pefloxacin plus ceftriaxone or pefloxacin plus amikacin. However, combinations
with ceftriaxone resulted in more frequent resistance to ceftriaxone than did
ceftriaxone alone. This was the case in P. aeruginosa infections treated with
ceftriaxone plus amikacin (p less than 0.01), and in S. marcescens infections
treated with ceftriaxone plus pefloxacin (p less than 0.05). Despite these
certain notable exceptions, our data confirm that in most cases combination
therapy does limit the emergence of resistance.
PMID: 3088999, UI: 86265701

6f. Am J Med 1983 Aug 29;75(2A):30-41 
Morphologic changes produced by amdinocillin alone and in combination with
beta-lactam antibiotics: in vitro and in vivo.
Kramer MJ, Mauriz YR, Timmes MD, Robertson TL, Cleeland R

Scanning electron microscopy was used to study the morphologic effects of
amdinocillin (mecillinam) when combined with several beta-lactam antibiotics in
vitro (Escherichia coli, three isolates; Klebsiella pneumoniae, one isolate)
and also in vivo (E. coli, one isolate). Ovoid forms were found in the cultures
of E. coli and K. pneumoniae following in vitro exposure to amdinocillin. This
characteristic in vitro effect was also produced in the amdinocillin-treated E.
coli-infected mouse. Varying degrees of filament formation were seen both in
vitro and in vivo with the other beta-lactam antibiotics tested. The in vitro
combination of amdinocillin with the beta-lactam antibiotics produced
morphologic effects on E. coli and K. pneumoniae (enhanced cell distortion and
lysis) not seen with the individual agents at the doses tested. Amdinocillin
was synergistic with ampicillin, carbenicillin, and cephalothin in mice
challenged with E. coli 736; scanning electron microscopy of bacteria from
peritoneal lavages of mice treated with these synergistic combinations
indicated that the organisms were more enlarged and distorted than those from
animals receiving the individual agents. The enhanced morphologic effect
observed in vivo was in agreement with the in vitro effect. Viable counts of
bacteria recovered from mice treated with ampicillin plus amdinocillin were
appreciably less than those from mice treated with each agent alone. The
morphologic results from the scanning electron microscopy study point to a
synergistic or enhanced effect of amdinocillin in combination with beta-lactam
antibiotics and are in accord with prior reports of the synergistic effects of
amdinocillin.
PMID: 6311003, UI: 83305802


7a. J Infect Dis 1995 Feb;171(2):385-92 
Modulation of nonspecific antimicrobial resistance of mice to Klebsiella
pneumoniae septicemia by liposome-encapsulated muramyl tripeptide
phosphatidylethanolamine and interferon-gamma alone or combined.
ten Hagen TL, van Vianen W, Bakker-Woudenberg IA
Department of Clinical Microbiology and Antimicrobial Therapy, Erasmus
University, Rotterdam, Netherlands.

Activation of the host defense system in a nonspecific way might provide tools
to support failing antibiotic treatment in certain infectious diseases. The
antimicrobial effect was investigated of liposome-encapsulated muramyl
tripeptide phosphatidylethanolamine (MTPPE) and interferon (IFN)-gamma and
liposome-coencapsulated MTPPE and IFN-gamma on Klebsiella pneumoniae septicemia
in mice. Prophylactic treatment of mice with five doses of liposomal MTPPE or
IFN-gamma increased survival from 0 to 65%. Administration of MTPPE and
IFN-gamma coencapsulated in liposome resulted in 100% survival. In vitro,
peritoneal macrophages by themselves were stimulated by these agents but were
unable to kill K. pneumoniae. However, production of both oxygen and nitrogen
intermediates increased when immunomodulators were added to macrophages. These
results indicate that effective prophylactic treatment of septicemia due to K.
pneumoniae with coencapsulated MTPPE and IFN-gamma is not solely due to
activation of the resident macrophages.
PMID: 7844375, UI: 95146786

7b. Ann Immunol (Paris) 1984 Jul-Aug;135D(1):59-69 
Immunological activities of RU-41740, a glycoproteic extract from Klebsiella
pneumoniae. I.--Activation of murine B cells and induction of interleukin-1
production by macrophages.
Guenounou M, Vacheron F, Zalisz R, Smets P, Agneray J

RU-41740, a glycoprotein extract from Klebsiella pneumoniae K2O1 strain, is an
immunomodulating compound which has been shown to reduce infectious episodes in
immunodeficient patients. Data from preliminary experimental designs suggested
that RU-41740 could affect several target cells, such as T cells, B cells and
macrophages. In the present report, we show that RU-41740 is a selective
B-lymphocyte activator. It induces blast transformation in Nude mouse spleen
cell cultures and in B-cell-enriched fractions obtained from normal mice. It
does not activate T lymphocytes to proliferate. Activation of mouse B
lymphocytes by RU-41740 is not affected by removal of adherent cells. RU-41740
also activates immunoglobulin secretion by murine B lymphocytes. Incubating
spleen cells from C3H/HeJ mice with RU-41740 results in cell proliferation and
activation of antibody-forming cells. This suggests that B-cell activation is
not due to LPS contamination. Other experiments show that RU-41740 can also
trigger mouse macrophages to produce interleukin-1 activity. Indeed,
supernatants from peritoneal adherent cells incubated in the presence of
RU-41740 can stimulate blastogenesis in thymocytes from C3H/HeJ mice. Thus,
B-cell activation and IL-1 production by macrophages could constitute two
additive mechanisms involved in immunomodulation induced by RU-41740.
PMID: 6385817, UI: 85021155

7c. J Clin Invest 1998 Oct 15;102(8):1583-90 
Antibacterial activity of human neutrophil defensins in experimental infections
in mice is accompanied by increased leukocyte accumulation.
Welling MM, Hiemstra PS, van den Barselaar MT, Paulusma-Annema A, Nibbering PH,
Pauwels EK, Calame W
Department of Radiology, Division of Nuclear Medicine, Leiden University
Medical Center, Leiden, The Netherlands. welling@rullf2.medfac.leidenuniv.nl

Neutrophil defensins (or human neutrophil peptides-HNP) are major constituents
of the azurophilic granules of human neutrophils and have been shown to display
broad-spectrum antimicrobial activity. Other activities of these defensins,
which are released from stimulated neutrophils, include cytotoxic, stimulatory,
and chemotactic activities toward a variety of target cells. We studied the
potential use of HNP-1 for antibacterial therapy of experimental bacterial
infections in mice. In experimental peritoneal Klebsiella pneumoniae infections
in mice, HNP-1 injection was shown to markedly reduce bacterial numbers in the
infected peritoneal cavity 24 h after infection. This antibacterial effect was
found to be associated with an increased influx of macrophages, granulocytes,
and lymphocytes into the peritoneal cavity. These leukocytes appeared to be a
requirement for the antibacterial effect, since in leukocytopenic mice
administration of HNP-1 did not display antibacterial activity. HNP-1 treatment
also reduced bacterial numbers in experimental K. pneumoniae or Staphylococcus
aureus thigh muscle infections. In this model, radiolabeled HNP-1 was found to
accumulate at the site of infection, whereas most of the injected HNP-1 was
rapidly removed from the circulation via renal excretion. These results
demonstrate that neutrophil defensins display marked in vivo antibacterial
activity in experimental infections in mice and that this activity appears to
be mediated, at least in part, by local leukocyte accumulation.
PMID: 9788972, UI: 99007396

7d. Chemotherapy 1998 May-Jun;44(3):149-52 
Antibacterial activity of peritoneal exudate in patients treated with 2 g
cefotiam for surgical anti-microbial prophylaxis.
Miglioli PA, Schoeffel U, Gabroska E, Allerberger F
Department of Pharmacology, University of Padua, Italy.

The objective of this study was to investigate the presence of antibacterial
activity in peritoneal exudate (PE) of patients treated with cefotiam (CFT).
CFT (2 g) was administered as a 'single-shot' antimicrobial prophylaxis to 6
patients at the beginning of colorectal resection. Samples of PE were collected
from each patient on days 1, 2 and 3 after surgery. CFT was detectable in the
samples of day 1 for 5 of the 6 patients. The influence of PE on antibacterial
activity of the antimicrobial drug was evaluated carrying out the MICs of CFT
against Escherichia coli K-12, E. coli (ATCC 10798), Klebsiella pneumoniae
(ATCC 1003), Proteus rettgeri (Sanelli) and Staphylococcus aureus (ATCC 29213)
with and without the addition of PE. The presence of PE enhanced the
antimicrobial activity of CFT against gram-negative strains, but not against S.
aureus (ATCC 29213). These results suggest the presence of substances in PE
that possess endogenous antibacterial activity. Thus, antimicrobial activity in
PE cannot be predicted by evaluating pathogen sensitivity in vitro only.
PMID: 9612603, UI: 98275550


8. Chung Hua Chieh Ho Ho Hu Hsi Tsa Chih 1993 Aug;16(4):205-8, 251 
[Histamine release from mast cell induced by main pathogenic bacteria of
respiratory infections in Guangzhou area].
[Article in Chinese]
Huang HL, Zhong NS
Guangzhou Institute of Respiratory Disease.

The methodology for isolating rat peritoneal mast cell was established. The
main pathogenic bacterial ultrasonicates of respiratory infection in GuangZhou
area have been examined for their ability to release histamine from purified
rat peritoneal mast cells. The results were as following: (1) 7 pathogenic
bacteria can cause direct release of histamine from rat mast cells. The most
effective histamine releasers were P. mira (histamine release percent is 26.89%
+/- 6.70%), Strep. Pneu (24.16% +/- 3.71%), E. Coli (20.40% +/- 4.83%) and K.
Pneu (17.91% +/- 4.60%). (2) Histamine release initiated by bacterial
ultrasonicates was completed within 30 minutes E. Coli differed from K. Pneu
induced histamine release was independent of its dose. (3) The products of K.
Pneu did not induce histamine release directly and bacterial hemolysin may play
a potent role in bacteria-induced release of histamine. Our findings provide
some evidence for mechanism by which respiratory bacterial infection can
precipitate or exacerbate attacks of bronchial asthma.
PMID: 7513619, UI: 94228615


9a. Antimicrob Agents Chemother 1993 Feb;37(2):276-80 
Effects of interleukin-8 on nonspecific resistance to infection in neutropenic
and normal mice.
Vogels MT, Lindley IJ, Curfs JH, Eling WM, van der Meer JW
Department of Medicine, University Hospital, Nijmegen, The Netherlands.

The effect of treatment with interleukin-8 (IL-8), a neutrophil-activating
cytokine, was investigated in normal and neutropenic mice infected with a
lethal dose of Pseudomonas aeruginosa, Klebsiella pneumoniae, or Plasmodium
berghei. Intraperitoneal (i.p.) IL-8 treatment was associated with accelerated
death when IL-8 was administered shortly before i.p. infection with P.
aeruginosa or shortly after i.p. infection with P. aeruginosa and K.
pneumoniae. Histopathological analyses demonstrated a tendency to more severe
organ lesions in IL-8-treated mice. Only nonneutropenic mice that received IL-8
shortly before the infectious challenge and at the site of infection were
protected by IL-8. Whether IL-8 is protective of or detrimental to the survival
of infection appeared to depend on the presence of bacteria at the injection
site and on the presence of neutropenia. IL-8 may be an important participant
in the cascade of interacting cytokines that is induced by the lethal
infectious challenge.
PMID: 8452358, UI: 93199301


9b. Zh Mikrobiol Epidemiol Immunobiol 1992 Feb;(2):62-4 
[Interferon type I in protective body reactions in an experimental Klebsiella
infection].
[Article in Russian]
Fil'chakov IV, Avdeeva LV, Anisimova IuN, Protsap EI, Spivak NIa

The study on mice with experimental generalized Klebsiella infection, carried
out with the use of microbiologic, immunologic and pathomorphologic methods,
revealed that the intraperitoneal injection of type I interferon into the
animals prevented their death and led to the rapid elimination of the infective
agent from their body, enhanced the phagocytic and metabolic activity of
polymorphonuclear lymphocytes of their peritoneal exudate, decreased the
manifestation of microcirculatory and dystrophic changes in the parenchyma of
their internal organs.
PMID: 1441818, UI: 93070692

9c. Microbiol Immunol 1990;34(2):185-95 
Protective effect of recombinant human interleukin-2 against lethal infection
caused by Klebsiella pneumoniae.
Iizawa Y, Nakao M, Kondo M, Yamazaki T
Research and Development Division, Takeda Chemical Industries, Ltd., Osaka.

The effects of recombinant human interleukin-2 (rIL-2) administered
prophylactically on the death of CBA/J mice challenged with Klebsiella
pneumoniae 27 intraperitoneally were examined. rIL-2 administered
subcutaneously at 20 micrograms per mouse for 7 days enhanced survival after a
lethal challenge. The injection of anti-asialo GM1 antibody did not influence
the effect of rIL-2. In mice given rIL-2, the number of peritoneal macrophages
increased, and the infiltration of polymorphonuclear leukocytes (PMN) into the
peritoneal cavity after the bacterial challenge was enhanced. In addition,
adoptive transfer of sera and peritoneal exudate cells (PEC), consisting of an
approximately equal number of macrophages and PMN, obtained from mice given
rIL-2 enhanced resistance to a K. pneumoniae infection, compared with adoptive
transfer of sera and PEC obtained from mice not given rIL-2. These results
indicate that rIL-2 protects mice from a lethal challenge with K. pneumoniae,
and suggest that the protective effect is due to an increase in the number of
phagocytic cells and in the cooperative activity of the sera and the phagocytic
cells.
PMID: 2189060, UI: 90265300


10. Microb Pathog 1992 Aug;13(2):145-55 
Modulation of surface antigen expression by Klebsiella pneumoniae in response
to growth environment.
Camprubi S, Smith MA, Tomas JM, Williams P
Department of Microbiology, Faculty of Biology, University of Barcelona, Spain.

Growth in pooled human body fluids [urine, serum and peritoneal dialysate (HPD)]
modulated the expression of cell envelope antigens in virulent (serotype O1:K1)
and avirulent (serotype O1:K66) Klebsiella pneumoniae strains. Marked variations
in the outer membrane protein (OMP) and lipopolysaccharide (LPS) profiles were
noted when broth-grown cells were compared with those of bacteria cultured in
body fluids. 
In particular, for the O1:K1 serotype strain, growth in the latter resulted in:
     (a) the expression of at least five iron-regulated OMPs in the 74-87
     kDa range, the pattern of which was medium dependent; 
     (b) alterations in the migration of the LPS core polysaccharide; and 
     (c) the reversion of isogenic O-:K+ and O-:K- mutants to the O+
     phenotype after growth in fresh serum but not in heat-inactivated
     serum, urine or HPD. 
Similar results were obtained for the O1:K66 serotype, although no variation in
the migration of the LPS core was noted. For both O1:K1 and O1:K66 serotypes,
neither the surface exposure of O1 serotype LPS nor the production of K-antigen
(capsular polysaccharide) was affected by growth in body fluids. No reversion
of K- mutants to the K+ phenotype was observed. 
     These data illustrate the phenotype flexibility of this opportunistic
     pathogen and emphasise the crucial role of the O- rather than the
     K-antigen in protecting K. pneumoniae from complement-mediated serum
     killing.
PMID: 1453927, UI: 93086384


11a. Antimicrob Agents Chemother 1988 Mar;32(3):364-8 
Influence of cephalosporins and iron on surface protein antigens of Klebsiella
pneumoniae in vivo.
Kadurugamuwa JL, Anwar H, Brown MR, Hengstler B, Kunz S, Zak O
          Division of Infectious Diseases, CIBA-GEIGY Ltd., Basel,
          Switzerland.
ab: The outer membrane protein (OMP) profiles of Klebsiella pneumoniae grown in
a rabbit peritonitis model in the presence or absence of cephalosporins were
investigated. Six high-molecular-weight OMPs (Mr 69,000 to 83,000) were induced
under iron-depleted conditions in vitro. Three of these proteins (the 69,000-Mr
protein [69K protein] and the 70K and 78K proteins) and trace amounts of the 73K
and 75K proteins were induced in the OM of bacteria infecting the peritoneal
cavity of rabbits. 

     Addition of iron either to the growth medium in vitro or to the
     peritoneum in vivo repressed the expression of these proteins.
     Cephaloridine had no significant effect on the OMP profiles. 

     An additional 56,000-Mr protein was observed in the OM of bacteria
     cultivated in vivo in the presence of CGP 17520 and also to a lesser
     extent in vivo under conditions of iron excess. 

A difference in recognition of OM antigens between cells grown in vitro and in
vivo was observed by immunoblotting techniques. The 26K, 27.5K, and 28.5K
antigens present in the OM of cells grown in vitro (but not in vivo) were
recognized by antibodies raised against bacteria cultivated in vitro under
conditions of iron depletion, but were not recognized by antisera raised against
bacteria harvested directly from infections. Antisera raised against a
nonencapsulated K. pneumoniae strain caused no agglutination of encapsulated K.
pneumoniae grown in vivo in the absence of cephalosporins. Rapid agglutination
was observed with this antiserum when the same encapsulated strain was grown in
vivo in the presence of either cephalosporin, indicating less occlusion of
critical antigens by the capsule.
PMID: 3284461, UI: 88208373

11b. Microbiol Immunol 1982;26(3):227-39 
Antimicrobial effect of human serum IgA.
Funakoshi S, Doi T, Nakajima T, Suyama T, Tokuda M

Serum IgA, IgG and colostrum secretory IgA prepared from specimens pooled from
a large number of human beings were shown to have measurable levels of
antibodies against Escherichia coli, Pseudomonas aeruginosa, Klebsiella
Pneumoniae, poliovirus, Coxsackie B virus, echovirus and influenza virus. Serum
IgA exerted a bacteriostatic effect in vitro on E. coli and P. aeruginosa,
which increased in the presence of the iron-binding proteins lactoferrin and
transferrin. This bacteriostasis was reduced when the iron-binding proteins
were saturated with iron. Similar results were obtained with IgG and secretory
IgA. The bacteriostatic effect of serum IgA was also shown in vivo, in the
peritoneal cavity of mice. The effect was suppressed by iron. Iron-chelating
substances, siderophores, excreted by E. coli diminished the co-operative
bacteriostatic effect of serum IgA and transferrin. Siderophore production by
E. coli was inhibited in the presence of serum IgA, but not when serum IgA was
deprived of specific antibody by absorption with E. coli. These results
indicate that serum IgA has a potent bacteriostatic effect in co-operation with
transferrin or lactoferrin because of the inhibitory effect of the specific
antibody on siderophore production by E. coli.
PMID: 6287178, UI: 82271515



12. Ann Inst Pasteur Immunol 1987 May-Jun;138(3):425-36 
RU-41740 (K. pneumoniae glycoprotein) enhances resistance to experimental
candidiasis and stimulates phagocytic functions.
Smets P, Salles MF, Rommain M, Zalisz R, Yagello M, Guenounou M
Centre de Recherches Immunologiques Roussel-UCLAF, Laboratoires Cassenne, Osny,
France.

RU-41740, a purified glycoprotein extract from Klebsiella pneumoniae, (which is
an efficient non-specific immune activator in a broad spectrum of in vitro and
in vivo reactions) was administered either orally or parenterally in the mouse.
It enhanced the resistance of mice to candidiasis, both in terms of survival
rate and a decrease in viable yeast cell recovery in kidneys. The drug
administered at 0.1 mg or 1 mg/kg augmented 4-fold the mean survival time (MST)
of animals infected with 1 to 2 X 10(6) Candida albicans, both by the
intraperitoneal and the intravenous route. The effect of the orally
administered drug was less striking but nonetheless present. At 10 mg/kg, the
MST of infected animals increased about 2-fold. In vitro, in the presence or
absence of zymosan, the drug at 10 or 100 micrograms/ml was able to stimulate
the phagocytic process of elicited mouse peritoneal cells (65%
polymorphonuclear cells, 35% macrophages) and human peripheral blood cells (95%
polymorphonuclear cells, 5% monocytes) in terms of activated oxygen species
production. The involvement of polymorphonuclear cells in the mechanisms of
natural resistance to C. albicans infection led us to discuss the role of these
cells as targets for the drug.
PMID: 3307833, UI: 88000143


13. Microbiol Immunol 1986;30(8):761-76 
Modification of susceptibility to Klebsiella pneumoniae during murine
cytomegalovirus infection.
Leung WC, Hashimoto K

The effect of murine cytomegalovirus (MCMV) infection on susceptibility to
bacterial infection was studied in mice by a combination of intraperitoneal
(ip) inoculation of a sublethal dose of MCMV with subsequent ip challenge of 2
X 10(3) cfu of a strain of Klebsiella pneumoniae (KP). When given alone, KP
produced a mortality of 30-40%. Mortality was increased when KP was given 1 to
7 days after MCMV injection with the peak increase at the 4th to 5th day when
100% mortality occurred. Virus levels in various organs of mice infected with
MCMV alone, or superinfected with KP did not differ. Bacterial counts on the
other hand, showed that increased mortality in mixed MCMV and KP infected mice
was due to an uncontrolled growth of bacteria at the site of primary lodgment,
i.e., the peritoneum, and severe systemic infection. Neutrophil response to
growth of KP during the first 3 days of bacterial infection was defective in
MCMV infected mice. While the initial clearance of KP from the blood was more
efficient in MCMV infected mice, probably due to activated reticuloendothelial
function, it did not protect the mice against KP infection. Using the in vivo
model, it was shown that poor neutrophil response and possibly other defective
neutrophil functions were responsible for increased mortality to KP infection
in MCMV infected mice.
PMID: 3023797, UI: 87063746

14a. Infect Immun 1981 Oct;34(1):166-70 
Specific lymphocyte blastogenic responses in children with cytomegalovirus and
herpes simplex virus infections acquired early in infancy.
Pass RF, Dworsky ME, Whitley RJ, August AM, Stagno S, Alford CA Jr

Cell-mediated immune responses in 27 infants and children with cytomegalovirus
(CMV) infection acquired between birth and 1 year of age were compared with
responses in 13 children who had neonatal herpes simplex virus (HSV) infection.
Infection was asymptomatic in 25 of 27 CMV-infected children; the 13 patients
with HSV infection were all ill as newborns. The median age when studied was 46
months for children infected with CMV and 24 months for those infected with HSV.
We measured lymphocyte transformation responses (LTRs) to CMV antigens in the
former group and to HSV type 1 (HSV-1) (and in six cases to HSV-2) in the latter
group, with the results expressed as a stimulation index. Based on the results
in seropositive and seronegative adult control subjects, stimulation indexes of
greater than or equal to 3 were considered indicative of a positive LTR. Among
the CMV-infected children, a positive LTR was observed in 0 to 13 assays
performed before 1 year of age, 3 of 8 assays performed between 1 and 4 years
of age, and 9 of 15 assays performed over 4 years of age. In contrast, a
positive LTR to HSV-1 was seen in 15 to 18 assays performed in children under
1 year of age and in 14 of 16 assays performed in survivors of neonatal HSV
infection older than 1 year. Six HSV-2-infected patients were tested
simultaneously 13 times with HSV-1 and HSV-2 antigens. Those patients under 6
months of age responded similarly to each antigen, whereas those who were older
had significantly higher LTRs to HSV-2. Children with CMV infection that was
acquired early had persistently diminished specific LTRs. In contrast, after
neonatal HSV infection, LTRs to HSV were present even in infancy and became more
specific for the infecting type with increasing age. 
PMID: 6271679, UI: 82052026


14b. Przegl Lek 1995;52(7):354-7 
[Congenital and acquired cytomegalovirus infection in infants confirmed by
virologic studies].
[Article in Polish]
Zawilinska B, Kruszewska M, Stopyrowa J, Zgorniak-Nowosielska I
Zakladu Wirusologii, Instytutu Mikrobiologii, Collegium Medicum, Uniwersytetu
Jagiellonskiego w Krakowie.

Sixty infants in whom clinical symptoms suspected of cytomegalovirus (CMV)
infection were studied. CMV infection was found in 50% of the subjects. The
diagnosis was based on studies of specific antibodies and isolation of the virus
from urine and/or throat swabs. In most of the children the examinations were
repeated several times, and clinical observations continued for 1 to 42 months
(avg. 18 months). IgM-class antibodies were detected in 26 children and in 18
the virus was isolated. In 3 infants, isolation of CMV virus was the only
evidence of active infection. Persisting viruria (avg. 11 months) and long-term
presence of Ig G antibodies, even to 44th month of life were also observed.
Congenital infection was diagnosed in 4 infants; the remaining ones acquired the
infection during the perinatal period or later. In 7 cases transfused blood
cannot be excluded as the source of infection. The clinical symptoms manifested
in infected and non-infected children were similar. There was a statistically
significant increase in the occurrence of hepatomegaly, splenomegaly,
hyperbiliru-binemia and diarrhoea in infected children. Congenital abnormalities
were found in 10 infected children, including 4 cases of congenital cytomegaly. 
PMID: 8525003, UI: 96106106


14c. Taylor-Wiedeman J.  Hayhurst GP.  Sissons JG.  Sinclair JH.
Polymorphonuclear cells are not sites of persistence of human
cytomegalovirus in healthy individuals.
          Journal of General Virology.  74 ( Pt 2):265-8, 1993 Feb.
  Polymorphonuclear leukocytes (PMNL) have been shown to harbour human
cytomegalovirus (HCMV) in viraemic patients, but to date PMNL of asymptomatic
healthy subjects have not been examined directly to determine whether this is
a normal site of HCMV persistence. Using the polymerase chain reaction (PCR),
paired DNA samples prepared from adherent peripheral blood mononuclear cells
(PBMC), which are known to be a site of persistence of HCMV, and PMNL of 10
healthy adults were analysed. All of seven individuals who were HCMV
seropositive, and one of three who were seronegative gave a reproducible signal
for HCMV DNA in their adherent PBMC, whereas none of the paired PMNL DNA samples
gave a positive result. The remaining two seronegative subjects showed no HCMV
DNA in either the PBMC or PMNL samples. In every case where PCR for HCMV was
negative, PCR amplification of a control human gene was used to show there was
no inability to amplify the DNA. We conclude that within the leukocyte
population of normal asymptomatic HCMV carriers, PMNL do not appear to harbour
persistent HCMV whereas adherent PBMC in the same subjects are a site of
persistence.

14d. Tokai J Exp Clin Med 1986;11 Suppl:59-63 
Experimental analysis of concurrent viral and bacterial infection in the mouse.
Hashimoto K
Department of Microbiology, School of Medicine, Tokai University, Kanagawa,
Japan.

The roles of both microorganisms in combined murine cytomegalovirus (MCMV) and
Klebsiella pneumoniae (KP) infection in the mouse, and host response to the
infection were investigated. Increased mortality in mixed MCMV and KP infected
mice was observed, and this was due to uncontrolled growth of bacteria in the
peritoneal cavity, and severe generalized infection. Neutrophil response to
growth of KP was defective in MCMV infected mice. While the initial clearance
of KP from the blood was more efficient in MCMV-infected mice, it did not
protect the mice against KP infection. In the in vitro experiments, the
neutrophil chemotaxis of MCMV-infected mice to KP was found to be lowered.
Tissue extract and serum of MCMV-infected mice exhibited chemotactic activity
for neutrophils, and, at the same time contained some factor(s) which reduced
chemotaxis of normal neutrophils to KP.
PMID: 2837837, UI: 88248936

15a. Microbiol Immunol 1979;23(5):369-82 
Effect of capsular polysaccharide of Klebsiella pneumoniae on host resistance
to bacterial infections. III. Further study of its effects on interactions
between peritoneal leukocytes and virulent Salmonella enteritidis.
Kato N, Kato O, Nakashima I, Naito S, Asai J

The mechanism for the infection-promoting effect of the capsular polysaccharide
of Klebsiella pneumoniae (CPS-K) was investigated using the experimental system
in which mice were infected intraperitoneally (i.p.) with a virulent strain of
Salmonella enteritidis immediately after i.p. injection of CPS-K. In the
peritoneal phagocytes of CPS-K-untreated control mice, approximately 70, 3, and
10% of phagocytized bacteria survived 6, 12, and 24 hr after challenge,
respectively, when calculated from the ratio of the number of cell-associated
viable bacteria, which was estimated by direct plate count, to the number of
phagocytized bacteria, which was estimated by microscopic observation of
stained smears. In contrast, almost all of the phagocytized bacteria were
viable throughout the experimental period in mice treated with CPS-K. The
electron microscopical findings of the phagocytes obtained 12 hr after
challenge showed that in the cells of mice treated with CPS-K almost all of the
phagocytized bacteria were morphologically intact, with some of them in the
stages of cell division, whereas in those of untreated control mice, almost all
of the phagocytized bacteria underwent digestive changes. When the reaction
product of acid phosphatase was examined by electron microscopy in the
phagocytes obtained 12 hr after challenge, the enzyme activity in the
phagosomes was very low in mice treated with CPS-K in comparison with that in
untreated control mice. Enzyme assays of the lysosomal and extralysosomal
fractions of peritoneal cells obtained at various times after challenge also
showed that release of acid phosphatase from the lysosomal fraction to the
extralysosomal fraction after bacterial challenge was inhibited in peritoneal
cells of mice treated with CPS-K.
PMID: 388155, UI: 80054148


15b. Jpn J Microbiol 1976 Oct;20(5):415-23 
Effect of capsular polysaccharide of Klebsiella pneumoniae on Host resistance
to bacterial infections. II. Effects on peritoneal leukocytes of normal mice
and mice infected with virulent Salmonella enteritidis.
Kato N, Kato O, Nakashima I

In normal mice, the total count of peritoneal leukocytes was markedly decreased
after intraperitoneal (i.p.) injection of the capsular polysaccharide of
Klebsiella pneumoniae (CPS-K) depending on the dosage injected. This decrease
was mainly due to the depletion of macrophages, and a decrease in the number of
lymphocytes occurred to a lesser extent. CPS-K in relatively smaller doses
mobilized polymorphonuclear neutrophilic leukocytes (PMN) into the peritoneal
fluid but it decreased them transiently in larger doses. In mice infected i.p.
with a virulent strain of Salmonella enteritidis, there was an abundant
emigration of PMN into the peritoneal fluid. When 200 mug of CPS-K was injected
i.p. immediately before bacterial challenge, emigration of PMN was markedly
delayed for 48 hr after infection. Associated with this suppressed emigration
of PMN, the numbers of macrophages and lymphocytes in the peritoneal fluid were
significantly less in mice treated with CPS-K than those in untreated control
mice for 48 hr after infection. The numbers of both cell-associated and
extracellular bacteria in the peritoneal fluid were markedly greater in mice
treated with CPS-K than those in untreated control mice. In both in vivo and in
vitro experiments, ingestion of bacteria by macrophages and PMN was not blocked
by CPS-K or neutral CPS-K, the active substance responsible for the
infection-promoting effect of CPS-K. It appeared that CPS-K somehow impaired
the intraphagocytic bactericidal activity.
PMID: 792533, UI: 77054454


15c. Jpn J Microbiol 1976 Jun;20(3):163-72 
Effect of capsular polysaccharide of Klebsiella pneumoniae on host resistance
to bacterial infections. I. Induction of increased susceptibility to infections
in mice.
Kato N, Kato O, Nakashima I

When Klebsiella pneumoniae capsular polysaccharide (CPS-K) from type 1, Kasuya
strain, was injected intraperitoneally (i.p.) immediately before i.p. bacterial
challenge, the survival time of mice infected with Salmonella enteritidis NUB
1 (virulent strain) was shortened and the mortality rate for mice infected with
S. enteritidis NUB 31 (avirulent strain) was enhanced. The promotion of
infection with S. enteritidis NUB 1 by CPS-K depended upon its dose, the effect
of CPS-K being demonstrable up to as little as 0.2 mug per mouse. In the case
of S. enteritidis NUB 31, the effect of CPS-K was detectable only when more
than 20 mug per mouse was injected. As a result of enumeration of bacterial
populations in the peritoneal washing, blood, liver and spleen, it was revealed
that CPS-K promoted in vivo growth of S. enteritidis NUB 1 and NUB 31. In
addition, CPS-K enhanced the mortality rate in mice infected with Streptococcus
pyogenes or Streptococcus pneumoniae. The peak CPS-K effect on infection with
S. enteritidis NUB 1 was seen when given immediately before bacterial
challenge. The active substance responsible for the infection-promoting effect
of CPS-K was neutral CPS-K, which is distinct from the O antigen and from
acidic CPS-K (the type-specific capsular antigen). Preparations of neutral
CPS-K isolated from the other three strains of K. pneumoniae exhibited a marked
infection-promoting effect comparable with that of preparations from the Kasuya
strain. Neutral CPS-K, with identical antigenicity to that from the Kasuya
strain, has already been found to exert a strong adjuvant effect on antibody
responses to various antigens in mice. No parallelism exists between
infection-promoting activity and adjuvant activity of neutral CPS-K.
PMID: 9528, UI: 77009621

16a. Infect Immun 1997 Apr;65(4):1546-9 
Binding of the type 3 fimbriae of Klebsiella pneumoniae to human endothelial
and urinary bladder cells.
Tarkkanen AM, Virkola R, Clegg S, Korhonen TK
Department of Biosciences, FIN-00014 University of Helsinki, Finland.

Binding of the two identified type 3 fimbrial variants of Klebsiella pneumoniae
to human endothelial EA-hy926 and bladder T24 cells was assessed. The
recombinant Escherichia coli strain LE392(pFK12), expressing plasmid-encoded
type 3 fimbriae of K. pneumoniae, adhered to both cell lines, and the fimbriae
purified from the strain bound to both cell lines in a dose-dependent manner.
Adhesiveness to both cell lines of chromosomally encoded type 3 fimbriae from
K. pneumoniae IApc35 was lower. No binding was detected with type 1 fimbriae of
K. pneumoniae. Both type 3 fimbrial variants exhibited a significantly lower
affinity for the cell lines than did S fimbriae of meningitis-associated E.
coli.
PMID: 9119502, UI: 97230338

16b. Infect Immun 1995 Oct;63(10):4046-53 
Activation of human endothelial cells by viable or heat-killed gram-negative
bacteria requires soluble CD14.
Noel RF Jr, Sato TT, Mendez C, Johnson MC, Pohlman TH
Department of Surgery, University of Washington School of Medicine, Seattle
98104, USA.

In response to bacterial lipopolysaccharides (LPS; endotoxin), endothelial
cells are converted to an activation phenotype expressing both proinflammatory
and procoagulant properties that include the induction of leukocyte adhesion
molecules and tissue factor expression. LPS-induced endothelial cell activation
requires a soluble form of the monocyte LPS receptor, sCD14. We evaluated the
capacity of multiple strains of gram-negative and gram-positive bacteria to
induce endothelial E-selectin and tissue factor expression through
sCD14-dependent pathways with cultured human umbilical vein endothelial cells
(HUVE). Both viable and heat-killed gram-negative bacteria (Bacteroides
fragilis, Enterobacter cloacae, Haemophilus influenzae, and Klebsiella
pneumoniae) but not viable or heat-killed gram-positive bacteria
(Staphylococcus aureus, Enterococcus faecalis, and Streptococcus pneumoniae)
induced prominent E-selectin surface expression detected by enzyme-linked
immunosorbent assay. Tissue factor activity on HUVE, indicated by factor X
activation, was induced in response to gram-negative bacteria but not in
response to gram-positive bacteria. Gram-negative bacteria induced
transcriptional activation in HUVE, indicated by the appearance of
E-selectin-specific mRNA and by the demonstration of activation of NF-kappa B,
a trans-activating factor necessary for E-selectin and tissue factor gene
transcription. In contrast, neither E-selectin mRNA nor activation of NF-kappa
B was detected in HUVE treated with gram-positive bacteria. Endothelial cell
activation by gram-negative bacteria in each of these assays was inhibited with
a monoclonal antibody (60bd) against CD14. Furthermore, CHO-K1 cells,
transfected with human recombinant CD14, responded to all strains of
gram-negative bacteria (viable or heat killed), indicated by CHO-K1 NF-kappa B
activation. We conclude that gram-negative bacteria induce endothelial cell
activation through a common sCD14-dependent pathway.
PMID: 7558318, UI: 96009765

17. A preliminary analysis of bbb infections and sequalae therefrom
HTTP://www.jorsm.com/~binstock/bbb-inf.htm