Biomedical subjects

J F Bohnsack

Publications and source records attributed to J F Bohnsack.

At least 55 records · Page 3Linked to original sources

PubMed1990

Human neutrophil adherence to laminin in vitro. Evidence for a distinct neutrophil integrin receptor for laminin.

We used mAbs against polymorphonuclear leukocyte (PMN) surface proteins to investigate the mechanisms by which stimulated human neutrophils (PMNs) adhere in vitro to laminin, the major glycoprotein of mammalian basement membrane. mAb IB4, which is directed against the common beta 2 chain of the CD11/CD18, only partially inhibited the adherence of PMA-stimulated PMNs to both laminin and to subendothelial matrices. In contrast, IB4 completely inhibited PMA-stimulated PMN adherence to gelatin, fibronectin, collagen IV, and endothelial cell monolayers. PMA-stimulated PMNs from a patient with severe congenital CD11/CD18 deficiency also adhered to laminin, but not to gelatin or endothelial cell monolayers. Therefore, PMA-stimulated PMNs adhere to laminin by both CD11/CD18-dependent and CD11/CD18-independent mechanisms. Expression of CD11/CD18-independent adherence to laminin was agonist dependent, occurring after stimulation with the calcium ionophore A23187 and recombinant TNF-alpha, but not with the chemotactic factors FMLP, platelet activating factor, or recombinant human C5a. Expression of CD11/CD18-independent adherence was also divalent cation dependent, occurring in the presence of Mg2+ but not Ca2+ as the sole added divalent cation. The mAbs AIIB2 and 13, which are directed against the beta 1 subunit of the VLA integrins, significantly inhibited the CD11/CD18-independent adherence of normal PMNs to laminin, and completely abolished the adherence of CD11/CD18-deficient PMNs to laminin. Both anti-beta 1 mAbs bound to PMNs, as demonstrated by flow cytometry, and immunoprecipitated a membrane molecule of Mr 130,000 daltons from 125I-labeled, detergent-solubilized PMNs. These data suggest that human PMNs possess beta 1 and beta 2 classes of integrins, and that both mediate PMN adherence.

Animals↗

PubMed1990

Effect of fibronectin on IgA-mediated uptake of type III group B streptococci by phagocytes.

Previous studies have shown that a type-specific IgA monoclonal antibody alone or in combination with fibronectin (Fn) enhances protective efficacy in two animal models of group B streptococcal infection. To investigate the mechanisms by which IgA mediates protection, the effects of Fn on phagocytosis of group B streptococci (GBS) opsonized with a type III-specific IgA monoclonal antibody were examined. Specific IgA alone or in combination with Fn did not promote the phagocytosis of GBS by polymorphonuclear leukocytes (PMNL). Fibronectin also had no significant effect on phagocytosis of IgA-opsonized GBS by monocytes. Specific IgA alone promoted phagocytosis of GBS by culture-derived macrophages in a dose-dependent fashion. Fibronectin enhanced macrophage uptake of the GBS opsonized in a suboptimal concentration of specific IgA (phagocytic index = 2.32 +/- 0.56 vs. 3.26 +/- 0.48 with Fn; P less than .05). These data suggest that protection against GBS in neonatal rats by a combination of Fn and specific IgA is mediated by macrophages rather than by PMNL or monocytes. Fibronectin may have a critical role in host defense at sites where IgA and macrophages predominate.

Antibodies, Monoclonal↗

PubMed1990

Mechanisms of tumor necrosis factor-alpha alteration of PMN adhesion and migration.

We have investigated the effects of recombinant human tumor necrosis factor-alpha (rhTNF alpha) on polymorphonuclear leukocytes (PMNs), concentrating on the mechanisms involved in the alterations of PMN-directed migration and adherence by this cytokine. RhTNF alpha profoundly suppressed PMN chemotaxis toward FMLP by 80%. At similar concentrations, it enhanced adhesion to gelatin-coated plastic dishes by more than tenfold and increased the expression of the CD11b antigen to 182% of the control. The monoclonal antibody 60.1, which is directed against the alpha chain of the CD11b/CD18 complex, completely blocked rhTNF alpha, induced inhibition of the chemotactic response to FMLP, and rhTNF alpha induced hyperadherence, suggesting that these effects were related to rhTNF alpha's effects on CD11b antigen expression. The fluid state of the PMN membrane was also decreased by rhTNF alpha. N-butanol, a known membrane fluidizer, partially inhibited the effect of rhTNF alpha on membrane fluidity and chemotaxis and completely reversed its effects on adherence and the expression of the CD11b antigen. Pentoxifylline, an agent that has previously been studied for its ability to prevent some effects of rhTNF alpha on PMNs, completely prevented the effect of rhTNF alpha on chemotaxis, the expression of the CD11b antigen, and membrane fluidity. Pentoxifylline partially prevented changes in adherence caused by this cytokine. Increased CD11b antigen expression caused by rhTNF alpha may result in enhanced PMN adhesion and suppression of migration. These events may, in turn, lead to the accumulation of PMNs on the vascular endothelium, resulting in the extensive vascular and tissue damage that is seen in gram-negative sepsis.

1-Butanol↗

PubMed1989

An IgA monoclonal antibody directed against type III antigen on group B streptococci acts as an opsonin.

We have investigated the mechanisms by which a murine IgA mAb directed against the type III Ag (IgA anti-III mAb) of group B streptococci (GBS) protects neonatal rats from lethal infection with these organisms. Purified IgA anti-III mAb enhanced phagocytosis of type III GBS by rat peritoneal macrophages in vitro by fourfold compared with phagocytosis of buffer-treated GBS. In the absence of antibody, neonatal rat serum did not promote phagocytosis, but addition of neonatal rat serum to GBS opsonized with IgA anti-III led to a sevenfold increase in phagocytosis. Heat inactivation of C destroyed the ability of neonatal rat serum to enhance phagocytosis in the presence of IgA. C3 deposition was observed when GBS coated with IgA anti-III mAb were incubated in untreated neonatal rat serum or in serum treated with Mg/EGTA. This latter observation suggested that C3 deposition occurred through activation of the alternative pathway. The control IgA mAb MOPC 315 did not enhance GBS ingestion or C3 deposition on GBS. Depletion of C in vivo by using cobra venom factor abolished the protective effect of IgA anti-III mAb in the neonatal rat model. These data suggest that the ability of this IgA to activate C further enhances its opsonic activity and may be essential for its protective effect in vivo.

Animals↗

PubMed1989

Neutrophil activating factor (NAF) induces polymorphonuclear leukocyte adherence to endothelial cells and to subendothelial matrix proteins.

Neutrophil activating factor is a polypeptide cytokine released from stimulated mononuclear phagocytes and endothelial cells. We found that neutrophil activating factor induced time- and concentration-dependent binding of human polymorphonuclear leukocytes to endothelial monolayers and subendothelial matrix proteins, via a mechanism that involves altered expression of the leukocyte CD11/CD18 glycoproteins. Thus, neutrophil activating factor is a third mediator, in addition to platelet-activating factor and endothelial leukocyte adhesion molecule 1, that is synthesized by activated endothelium and that can induce polymorphonuclear leukocyte adhesion to endothelial cells. Because NAF is released into the pericellular fluid, it may also stimulate binding of the leukocytes to exposed subendothelial structures at sites of vascular injury.

Antigens, Surface↗

PubMed1989

Effect of immune globulin intravenous on opsonization of bacteria by classic and alternative complement pathways in premature serum.

The mechanisms involved in the effect of immune globulin intravenous (IGIV) on bacterial opsonization by both complement pathways in premature serum were elucidated in this study. Of the bacteria used, Staphylococcus aureus and Salmonella enteritidis were nonencapsulated while Streptococcus pyogenes and Escherichia coli 07 K1 were encapsulated. As demonstrated by indirect immunofluorescence, IGIV showed specific antibody titers of 1:32 for S. aureus and S. enteritidis and of 1:8 for S. pyogenes and E. coli 07 K1. IGIV alone had no direct opsonic activity against these organisms. Addition of IGIV did not alter the opsonic activity of normal adult serum against these organisms. In contrast, addition of IGIV promoted the opsonic activity of premature serum against the nonencapsulated bacteria to levels matching that in normal adult serum. The IGIV preparation significantly improved the opsonization of bacteria by the classic (from 39 to 68% of that in adult serum) and alternative (from 22 to 97% of that in adult serum) complement pathways in premature serum. IGIV also markedly augmented C3 deposition on the bacteria by both complement pathways. These studies suggest that IGIV containing high titers of specific antibodies promote opsonization of bacteria by the enhancement of complement pathway activation, especially the alternative pathway, in premature serum.

Complement System Proteins↗

PubMed1988

Group B streptococci inhibit the chemotactic activity of the fifth component of complement.

Infection with group B streptococci (GBS) is associated with a poor acute inflammatory response in which neutrophils fail to localize at the site of invasion. In the present studies, we have examined the effects of group B streptococci on C-derived chemotactic activity in human serum. Fresh human serum was activated to form C5a and C5adesarg by incubation with zymosan. The activated serum was then incubated with group B organisms, centrifuged, and the supernatants tested for chemotactic activity for human polymorphonuclear leukocytes. Group B organisms caused a dose-dependent decrease in C-dependent chemotactic activity. The degree of inhibition was profound with 1 X 10(9) bacteria/ml (10% of control). Experiments indicated that significant chemotactic factor inactivation occurred within 2 min of exposure to GBS organisms, while maximal inhibition occurred after 30 min incubation. A number of different strains of GBS of types I, II, and III possessed inhibitory activity. In contrast, group D streptococci, Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae failed to inhibit the C-derived chemotactic activity in human serum. Group A streptococci that were M protein positive also inactivated C-dependent chemotactic activity in serum, as previously reported. The inhibitory activity of the GBS strains could be abolished by heat or trypsin treatment but not by neuraminidase, pronase, or pepsin. C5a levels in zymosan-activated serum as measured by RIA were not decreased after incubation with an inhibitory strain suggesting that absorption was not involved. SDS-PAGE analysis revealed that group B streptococci degrade the C5a molecule, increasing its electrophoretic mobility by removing a fragment with a m.w. of approximately 650 Da. Thus, one of the reasons for the poor inflammatory response at the site of GBS infection may reside in the ability of these pathogens to inactivate C-derived inflammatory mediators. The GBS C5a-ase activity probably serves as an additional virulence factor for these organisms contributing to the poor inflammatory response characteristic of group B streptococcal infection.

Aminopeptidases↗

PubMed1988

CR2 ligands modulate human B cell activation.

A considerable body of evidence from this and other laboratories indicates that complement receptor type 2 (CR2) modulates B cell activation and growth. In the present studies we have examined the effects of three different types of CR2 ligands, i.e., monomeric, aggregated, and latex-bound C3dg; mAb to different CR2 epitopes; and UV-inactivated, non-transforming EBV (EBVUV) for their actions on highly purified, high density resting tonsil B cells. Although none of these ligands induced B cells to enter the cell cycle or synergized with either anti-mu or low m.w. B cell growth factor in triggering B cell mitogenesis, aggregated C3dg, latex-bound C3dg, the OKB7 anti-CR2 mAb, and EBVUV-enhanced thymidine incorporation by phorbol ester-activated tonsil B cells. Such enhancement was not T cell or monocyte dependent. The major action of the CR2 ligands thus seems to be to enhance the transition of B cells activated by certain stimuli from the G1 to the S phase of the cell cycle. In contrast to the action of aggregated and latex-bound C3dg, monomeric C3dg was inhibitory for phorbol ester and aggregated C3dg-induced B cell activation. The HB-5 anti-CR2 mAb, which reacts with a different epitope on CR2 from that of OKB7, did not synergize with PMA in B cell activation. These data provide additional evidence for a role for the CR2 in the control of B cell growth and provide a useful model for studying the CR2-mediated signals that affect the growth of B cells.

Adjuvants, Immunologic↗

PubMed1988

Effects of fibronectin on the interaction of polymorphonuclear leukocytes with unopsonized and antibody-opsonized bacteria.

Fibronectin (Fn) affects the interaction of polymorphonuclear leukocytes (PMNLs) with certain bacteria. Fn alone enhanced the response, in a chemiluminescence (CL) assay, of PMNLs to Staphylococcus aureus (P less than .05) and Staphylococcus epidermidis (P less than .01) but had no effect on type III, group B streptococci (GBS) or Escherichia coli. When GBS or E. coli were first preopsonized in antibody, Fn significantly enhanced the CL response of PMNLs (P less than .05). The intracellular metabolic inhibitor NaN3 but not the extracellular scavengers superoxide dismutase or human serum albumin inhibited Fn-enhanced CL; this fact suggests that enhancement of the respiratory burst by Fn is an intracellular event. We used an acridine orange-crystal violet monolayer assay to examine the effects of Fn on ingestion and intracellular killing of bacteria by PMNLs. Fn alone promoted uptake and killing of S. aureus (P less than .01) and S. epidermidis (P less than .05) by PMNLs but did not enhance monolayer phagocytosis of GBS or E. coli, unless these bacteria were preopsonized in antibody (P less than .01).

Azides↗

PubMed1988

Mechanism of inhibition of immunoglobulin G-mediated phagocytosis by monoclonal antibodies that recognize the Mac-1 antigen.

We have investigated the effects of the monoclonal antibodies against the cell surface molecule Mac-1 on C3bi-mediated rosetting and IgG-mediated rosetting and phagocytosis by human peripheral blood monocytes. Highly purified M1/70 F(ab')2, used in the fluid phase, inhibited both monocyte functions. Half-maximal C3bi rosette inhibition occurred at a concentration of 2 nM F(ab')2 M1/70. An equivalent decrease in IgG-mediated rosetting required 10 nM M1/70 F(ab')2, and 50% inhibition of IgG-mediated phagocytosis required 7 nM antibody. Mo-1 F(ab')2 inhibited EC3bi binding with an ID50 of 0.3 nM, whereas 50% decrease in IgG-mediated rosetting required 70 nM of this antibody. OKM1 did not inhibit rosettes of sheep erythrocytes opsonized with IgG antibody (EA) at all. F(ab')2 M1/70 did not affect the binding of monomeric human IgG to monocytes, but did substantially decrease the binding of IgG aggregates. Half-maximal inhibition of aggregated IgG binding at 0 degrees C occurred at 8 nM F(ab')2 M1/70, very close to the concentration that caused equivalent inhibition of IgG-mediated phagocytosis. Aggregated IgG inhibited the binding of radiolabeled M1/70 to monocytes by approximately 40%, suggesting that some, but not all Mac-1 molecules were associated with IgG receptors under these conditions. When cells were allowed to adhere to surfaces coated with M1/70 or Mo-1 F(ab')2, C3bi-mediated rosetting was inhibited, but IgG mediated-phagocytosis was unaffected. Moreover, the dose response of inhibition of phagocytosis by fluid-phase F(ab')2, of anti-Mac-1 monoclonals was similar on monocytes adherent to albumin-coated and antibody-coated surfaces. Kinetic experiments showed that even prolonged incubation of monocytes on M1/70 coated surfaces did not lead to inhibition of EA binding nor did these incubations alter the dose response for inhibition of EA binding by fluid-phase M1/70 F(ab')2. This suggested that not all molecules recognized by M1/70 are freely mobile in the plasma membrane. Indeed, only approximately 60% of 125I-M1/70-biding sites were lost even after 4 h when monocytes were adherent to M1/70-coated surfaces. We conclude that some anti-Mac-1 antibodies can inhibit EA binding because of their epitope specificity, independent of any direct interaction with monocyte Fc receptors. This interference with IgG-Fc receptor-mediated binding and ingestion apparently occurs because of antibody binding to a subpopulation of Mac-1 molecules which are associated with IgG Fc receptors and remain on the apical membrane of monocytes adherent to anti-Mac-1-coated surfaces. We suggest that there may be two functionally distinct molecules on human monocytes recognized by M1/70 and Mo-1 that can be distinguished by their mobility in the plane of the monocyte membrane. The more mobile form of Mac-1 is involved in C3bi rosettes, and does not affect IgG-mediated phagocytosis. The other antigen recognized by M1/70 does not diffuse within the plane of the membrane; ligation of the latter molecule by antibody is associated with inhibition of IgG-mediated phagocytosis.

Antibodies, Monoclonal↗

PubMed1987

Evidence for a conformational change in human fibronectin which occurs between 4 and 37 degrees C.

The effects of temperatures between 4 and 37 degrees C on the structure of human fibronectin (Fn) using monoclonal antibodies (MAb) with specificity for Fn have been studied. Three MAb bound to Fn in solution better at 4 than at 37 degrees C, assessed by both ELISA and immunoprecipitation. These MAb detected temp-dependent conformational changes in Fn which were not associated with a major refolding of the Fn molecule, since the S20,w of Fn and the binding of Fn to polyclonal anti-Fn were unchanged by temp shifts from 4 to 37 degrees C. Fn also was adhered directly to ELISA plates at 4 and at 37 degrees C, and then probed with MAb, with detergents, and with trypsin. One MAb bound better to surface adherent Fn when the Fn had bound to the surface at 4 rather than 37 degrees C. Both SDS and polyoxyethylene (20) sorbitan monolaurate (Tween-20) caused the release of more adherent Fn from surfaces to which it had adhered at 4 degrees C. Finally, trypsin released Fn fragments more rapidly and more completely from surfaces coated at 4 than at 37 degrees C. It is concluded that Fn has different conformations at 4 and 37 degrees C in solution and after adherence to surfaces, as reflected by MAb binding, detergent sensitivity and protease susceptibility. This conformational difference is not associated with alterations in the physiochemical properties of the molecule and is thus a result of a less extensive intramolecular rearrangement than the conformational changes resulting from manipulations of ionic strength or pH, and from binding to collagen or hydrophobic surfaces. Nonetheless, the more subtle conformational change demonstrated here may be important for the biological properties of Fn.

Antibodies, Monoclonal↗

PubMed1987

Demonstration of defective C3-receptor-mediated clearance by the reticuloendothelial system in patients with acquired immunodeficiency syndrome.

The function of macrophage C3 receptors was assessed in vivo by measuring the clearance of C3-sensitized autologous erythrocytes in seven acquired immunodeficiency syndrome (AIDS) patients, eight healthy homosexual men, eight healthy heterosexual men, and four infected controls. Healthy heterosexual men had an initial clearance of 50.1 +/- 2.0% of the inoculum, with a release of a small portion of these cells (10.9 +/- 1.3%) into the circulation. Healthy homosexual men had a greater initial clearance of 66.0 +/- 4.2% (P less than 0.01) followed by a similar release (14.0 +/- 3.3%). AIDS patients had an initial clearance of 60.6 +/- 7.5% but had a relatively large release of cells (25.6 +/- 3.2%) (P less than 0.005 vs. heterosexuals; P less than 0.05 vs. homosexuals), suggesting a failure of macrophage phagocytosis. Infected controls had an initial clearance of 59.4 +/- 4.9%, with a release of 19.6 +/- 3.8% (P = NS vs. AIDS). These data, in addition to Fc-receptor dysfunction, demonstrate a global reticuloendothelial system dysfunction in AIDS patients. This may contribute to their frequent infections with opportunistic pathogens and inappropriate immune responses against these microorganisms.

Acquired Immunodeficiency Syndrome↗

PubMed1986

Interaction of culture-derived macrophages with the fibroblast-binding domain of fibronectin is a necessary but inefficient signal for fibronectin enhancement of CR1-mediated phagocytosis.

Human plasma fibronectin (Fn) enhances ingestion of opsonized particles through its interaction with phagocytic cells. To better characterize the site or sites on Fn responsible for this effect, we subjected Fn to limited proteolytic cleavage by chymotrypsin and used affinity and gel filtration chromatography to isolate a 110,000 dalton cell-binding fragment, a 60,000 dalton fragment that bound both heparin and gelatin, and 50,000 and 45,000 dalton fragments that bound to gelatin but not heparin. The cell-binding fragment mediated adhesion and spreading of fibroblasts on glass slides, whereas the heparin-gelatin and gelatin-binding fragments failed to cause fibroblast spreading. At high concentrations, the cell-binding fragment doubled phagocytosis of C4b-coated sheep erythrocytes by human monocyte-derived macrophages, whereas equal concentrations of the other fragments had minimal enhancing effect on phagocytosis. Interestingly, the effect of the cell-binding fragment on CR1-mediated phagocytosis was always less than the effect of intact Fn, even when a 40-fold higher molar concentration of the cell-binding fragment was used. Fab of a monoclonal anti-Fn, HFn 7.1, which recognizes the 110,000 dalton cell-binding fragment of Fn and inhibits fibroblast binding, blocked enhancement of CR1-mediated phagocytosis by intact Fn. Fab of Fn 8, a monoclonal anti-Fn that binds the heparin-gelatin-binding fragment, failed to inhibit the Fn effect. These data suggest that interaction of the macrophage with the cell-binding domain of Fn is critical for the Fn effect on CR1-mediated phagocytosis. In addition, there may be other domains of the Fn molecule that have a role in augmenting the Fn-phagocyte interaction.

Antibodies, Monoclonal↗

PubMed1986

The role of the spleen in resistance to infection.

The spleen serves two major critical functions in protecting against bloodstream bacterial infections--it is a phagocytic filter that removes bacteria from the bloodstream and it is an antibody-producing organ. Although the liver appears to remove the majority of well-opsonized bacteria from the bloodstream, the spleen plays an important role through its ability to sequester bacteria that are not as well opsonized and, thus, is of critical importance in the nonimmune host. The spleen may also be critical in the production of opsonizing antibodies, which are important for the rapid and efficient removal of bacteria from the bloodstream.

Adult↗

PubMed1985

Connective tissue proteins and phagocytic cell function. Laminin enhances complement and Fc-mediated phagocytosis by cultured human macrophages.

Brief exposure of culture-derived human macrophages to laminin, a glycoprotein component of all mammalian basement membranes that has a molecular weight of 1,000,000, led to enhancement of subsequent macrophage phagocytosis of EAC4b, EAC3bi, and EAIgG (sheep erythrocytes sensitized with IgG anti-Forssman antibody). This effect on macrophage phagocytosis occurred with both substrate-adherent and fluid phase laminin. Preincubation of macrophages, but not of EAC4b, with laminin led to augmentation of phagocytosis, suggesting that interaction with the phagocytic cell, but not with the opsonized particle, was required for laminin's effect. Laminin-stimulated phagocytosis of EAC4b was blocked entirely by a monoclonal antibody to CR1. Direct comparison of the phagocytic ability of macrophages adherent to laminin- and fibronectin-coated glass slides showed that fibronectin had a somewhat greater enhancing effect on phagocytosis. Nonetheless, the phagocytosis-enhancing effect of laminin was not due to contamination of the purified laminin preparation by fibronectin, since the laminin preparation was free of fibronectin, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and enzyme-linked immunosorbent assay; in addition, laminin-enhanced phagocytosis was decreased in the presence of laminin-specific antibodies. Laminin inhibited macrophage adherence and spreading, but selection of a laminin-binding macrophage subpopulation could not account for the laminin-induced increases in phagocytosis. We hypothesize that interaction with extracellular matrix proteins may represent an important activation stimulus both to the macrophages normally present in the extravascular compartment and to the phagocytic cells that have emigrated from the blood-stream into areas of inflammation.

Adjuvants, Immunologic↗

PubMed1985

The C1q subunit of the first component of complement binds to laminin: a mechanism for the deposition and retention of immune complexes in basement membrane.

The C1q subunit of complement component C1 is known to bind to immune complexes, which often are deposited in basement membrane. We investigated the possibility that this deposition is a result of binding to laminin, a large basement membrane glycoprotein. C1q showed saturable binding to immobilized laminin; this binding was increased at reduced ionic strength. Intact C1 did not bind laminin. A ternary complex was formed by laminin, C1q, and aggregated IgG. This complex formation was dependent on and proportional to the amount of C1q bound to the aggregated IgG. Binding of laminin to C1q occurred with a Kd of 2 nM and was stronger than the binding of C1q to fibronectin. Preliminary data, including electron micrographs of rotary-shadowed preparations, suggest that laminin binds to the collagen-like tail of C1q. Electron microscopy localized the site of interaction with C1q to a short arm of laminin. Since laminin is found only in basement membranes, the interaction between laminin and C1q could be involved in the deposition and retention of immune complexes in these structures.

Antigen-Antibody Complex↗

PubMed1985

Fibronectin-enhanced phagocytosis of an alternative pathway activator by human culture-derived macrophages is mediated by the C4b/C3b complement receptor (CR1).

We examined the ability of human monocytes and culture-derived macrophages under serum-free conditions to phagocytose desialated sheep erythrocytes (E), an activator of the alternative pathway of human complement. Freshly derived monocytes ingested desialated erythrocytes, but the degree of phagocytosis varied among individual donors. However, exposing the phagocyte to intact plasma fibronectin (Fn) had no effect on monocyte phagocytosis. Macrophages derived from monocytes in culture were far more efficient at ingesting desialated E, and the extent of phagocytosis was proportional to the degree of desialation. Although exposure of macrophages to substrate-bound Fn or fluid-phase Fn enhanced the phagocytosis of desialated E, pretreatment of desialated E with Fn did not enhance phagocytosis, demonstrating that Fn acted through an interaction with the macrophages. Fn-enhanced phagocytosis of desialated E was inhibited by treating macrophages with a monoclonal antibody to the C4b/C3b receptor (CR1), but not with a monoclonal antibody to the receptor for C3bi (CR3). Addition of cobra venom factor (CVF) to the macrophages also inhibited Fn-enhanced phagocytosis of desialated E. Phagocytosis of IgG-sensitized E, either in the absence or in the presence of Fn, was not significantly affected by anti-CR1 or CVF, demonstrating that these reagents did not lead to a general inhibition of phagocytosis. These experiments suggest that macrophages may deposit enough C3b onto desialated E to cause CR1-mediated phagocytosis in the presence of Fn. The ability of macrophages to opsonize and ingest foreign particles that activate complement may be critically important in areas of inflammation where concentrations of serum-derived specific opsonins may be inadequate.

Animals↗

PubMedDate not supplied

Bacterial genetics and human immunity to group B streptococci.

Serotype III group B Streptococcus agalactiae (GBS) are the most common cause of neonatal sepsis and meningitis. We have classified type III GBS by restriction digest patterns of chromosomal DNA and demonstrated that a subgroup of genetically related strains (RDP type III-3) causes the majority of type III GBS neonatal infection. Genetic differences between type III GBS strains contribute significantly to differences in virulence and host immune responses. While 100% of less virulent RDP type III-1 and III-2 organisms express C5a-ase, an inhibitor of neutrophil chemotaxis, only 63% of virulent RDP type III-3 isolates have functional C5a-ase. Functional differences in type III GBS C5a-ase are attributable to a shared genetic polymorphism, supporting our genetic classification. The mean capsular sialic acid content of virulent RDP type III-3 strains is significantly higher than that of less virulent strains, suggesting that capsular sialylation is also genetically regulated. C5a-ase is not critical for all RDP type III-3 strains to be invasive because the higher capsular sialic acid content of III-3 strains limits complement activation. The identification of these and additional genetic differences between GBS strains has important implications for our understanding of the pathogenesis of these important human infections.

Adhesins, Bacterial↗