Biomedical subjects
J A Winkelstein
Publications and source records attributed to J A Winkelstein.
Bacterial lipoteichoic acid sensitizes host cells for destruction by autologous complement.
Lipoteichoic acids (LTA) released by gram-positive bacteria can spontaneously bind to mammalian cell surfaces. In the present study, erythrocytes (E) sensitized with pneumococcal LTA (LTA-E) were used as a model system to determine if LTA could render host cells susceptible to damage by autologous complement. Complement (C)-mediated lysis of LTA-E from normal rats and normal humans occurred when these cells were incubated in their respective autologous sera in vitro. In addition, when LTA-E from a C2-deficient human and from C4-deficient guinea pigs were incubated in their autologous sera, there was significant lysis in vitro, demonstrating a role for the alternative pathway. The in vivo survival of 51Cr-labeled autologous LTA-E was also studied. Only 2.9% of autologous LTA-E remained in the circulation of normal rats after 90 min. In contrast, 31.2% of autologous LTA-E remained in the circulation of rats depleted of C3. Intravascular hemolysis accounted for the clearance of LTA-E in the normal rats, whereas liver sequestration was responsible for clearance in the C3-depleted rats. These results demonstrate that LTA can render the host's cells susceptible to damage by its own complement system, establishing this as a possible mechanism of tissue damage in natural bacterial infections.
The clinical manifestations of a genetically determined deficiency of the third component of complement in the dog.
The clinical manifestations of a genetically determined deficiency of C3 were examined in a closed colony of dogs. One hundred and twelve dogs, including twenty C3-deficient dogs, were studied over a period of 6 years. Five of the C3-deficient dogs developed significant bacterial infections, such as pneumonia, sepsis, and pyometra, which were caused by Clostridium spp., Escherichia coli, and Klebsiella spp. Two of the C3-deficient dogs who had had significant infections also subsequently developed renal disease. Secondary amyloidosis was the predominant renal lesion in one dog. The predominant renal lesion in the second dog was membranoproliferative glomerulonephritis, although some amyloid was also present. The two dogs with renal disease also had positive rheumatoid factors. No other clinical or serological evidence of autoimmune disease or immune complex disease has been found. None of the dogs heterozygous for C3 deficiency, and none of the homozygous normal dogs in the colony has developed significant bacterial infections or renal disease. Thus, dogs deficient in C3, like C3-deficient humans, demonstrate both an increased susceptibility to infection and renal disease.
Purification of the third component of canine complement.
A rapid, simple procedure has been developed for the purification of the third component (C3) of canine complement. Dog plasma was initially fractionated by precipitation with 4% (w/v) polyethylene glycol (PEG) 4000. The supernatant was depleted of plasminogen using a Sepharose 4B-lysine column, and the effluent was again fractionated with PEG 4000 at 16% (w/v). The precipitate was resuspended and passed over a DEAE-Sephacel column. The fractions containing hemolytically active C3 were pooled, concentrated, and passed over a CM-Sepharose CL-6B column to yield the final purified product. Rabbit anti-whole dog serum identified only one protein in the purified material on immunoelectrophoresis. When injected into a rabbit, the purified product raised an antisera which reacted with only one protein in both whole dog serum and the purified product. Analysis by SDS-PAGE revealed a single band of MW 179,000 +/- 7,000 (+/- 1 S.D.) daltons which, upon reduction with 2-mercaptoethanol, resulted in 2 bands of 114,000 +/- 6,000 daltons and 65,000 +/- 3,500 daltons. Final recovery was 18% with respect to C3 antigen and 9% with respect to C3 hemolytic activity.
Activation of the alternative complement pathway by pneumococcal lipoteichoic acid.
Cell wall teichoic acids of some gram-positive bacteria are potent activators of the alternative pathway of complement. It is unclear, however, whether the other form of teichoic acid, cell membrane lipoteichoic acid (LTA), can also activate the alternative pathway. In the present study, radiolabelled pneumococcal LTA was found to bind spontaneously to sheep erythrocytes in a temperature- and time-dependent fashion. In addition, the presence of pneumococcal LTA on the erythrocyte surface was verified by the fact that they could be agglutinated by a myeloma protein (TEPC-15) specific for choline, a constituent of pneumococcal LTA. Pneumococcal LTA when fixed to the surface of erythrocytes was able to activate the alternative pathway of complement in both guinea pig serum deficient in the fourth component of complement and human serum deficient in the second component of complement, resulting in lysis of the sensitized erythrocytes. The sensitizing principle of the LTA preparation was removed before erythrocyte sensitization by immunoabsorption, using the choline-specific TEPC-15 myeloma protein. These data demonstrate that purified pneumococcal LTA will bind to sheep erythrocytes and endow them with the ability to activate the alternative pathway.
Oral administration of human serum immunoglobulin in immunodeficient patients with viral gastroenteritis. A pharmacokinetic and functional analysis.
We examined the pharmacokinetics and immunological activity of human serum immunoglobulins (HSG) possessing anti-rota-virus activity which were orally administered to three children with primary immunodeficiency syndromes and prolonged gastrointestinal excretion of rotavirus. Detailed analysis of the excretion of immunoglobulins labeled with biotin or I125 revealed that approximately 50% of the recovered radioactivity was excreted in the stools over a 3-d period. Approximately half of the excreted radioactivity recovered in the stool was in a macromolecular form with immunological activity. The remainder of the recovered radioactivity was excreted in the urine as low molecular weight fragments or free iodide. In addition, immunological and chromatographic analyses revealed that the oral administration of HSG resulted in the generation of rotavirus-specific immune complexes in the gastrointestinal tract with a subsequent decrease in the presence of uncomplexed rotavirus antigen. These studies indicate that orally administered HSG can survive passage in the gastrointestinal tract in an immunologically active form, and that the oral administration of immunoglobulins with specific reactivities has potential for the prevention or treatment of gastrointestinal infections.
X-linked agammaglobulinemia: an analysis of 96 patients.
We have defined the clinical presentation and course of X-linked agammaglobulinemia (X-LA) by means of a multi-center retrospective survey of 96 patients. Infections were the most common presenting feature of patients with X-LA. The most frequent infections involved the upper respiratory tract (75%), lower respiratory tract (65%), gastrointestinal tract (35%), skin (28%), and central nervous system (16%). Clinical clues to the diagnosis of X-LA were the chronic or recurrent nature of infections, a family history of immunodeficiency, and infections at more than one anatomic location. Infections remained a significant problem after the diagnosis of X-LA was made and gamma-globulin prophylaxis had been instituted. One or more chronic infectious diseases occurred in 71% of patients. The respiratory tract was the most common site of disease, and the gastrointestinal tract was relatively spared. Patients died at a mean age of 17 years. The two major causes of death were chronic pulmonary disease with resultant cardiac failure, and disseminated viral infections which characteristically caused a dermatomyositis-like syndrome, hepatitis, pneumonitis, and meningoencephalitis.
Genetically determined variation in the complement system: relationship to disease.
Explore the source record for details and available documents.
Complement and the host's defense against the pneumococcus.
The complement system plays a critical role in resistance to Streptococcus pneumoniae. In the last two decades, a great deal of new knowledge has been generated on both the complement system and on the pneumococcus. Not only has this new knowledge helped produce a better understanding of how the complement system serves the host in its defense against S. pneumoniae, but in a more general fashion, the lessons learned from the interaction of complement and the pneumococcus have also provided insight into how the complement system functions in the defense of the host against a wide variety of other microorganisms. The following review will attempt to summarize current knowledge about the complement system, how it interacts at the molecular level with the pneumococcus, the biologic consequences of that interaction, and their significance in pneumococcal infections.
Pneumococcal-induced pulmonary leukostasis and hemodynamic changes: role of complement and granulocytes.
In the present study we investigated the cardiopulmonary dysfunction caused by systemic pneumococcal (PNC) disease. We studied the hemodynamic and hematologic effects of intravascular challenge with nonviable PNC in normal, granulocyte-depleted, and genetically C3-deficient dogs. In normal dogs PNC administration caused a decrease in cardiac output (CO) of 58% (p less than 0.001), an increase in pulmonary vascular resistance (PVR) of 151% (p less than 0.02), and an increase in systemic vascular resistance (SVR) of 72% (p less than 0.02). The PNC challenge also caused significant decreases in both circulating granulocytes (-73%; p less than 0.02) and platelets (-58%; p less than 0.02). Histologic examination revealed granulocyte plugging within small pulmonary vessels. PNC challenge in granulocyte-depleted and C3-deficient dogs resulted in hemodynamic and hematologic changes that were not significantly different from those seen in normal PNC-challenged animals. Infusion of PNC-activated plasma resulted in hematologic changes that were not significantly different from those seen in PNC-challenged animals. Infusion of plasma treated with ethylenediamenetetraacetic acid (EDTA) prior to PNC incubation induced the same hematologic alterations but the hemodynamic response was less pronounced. To control for PNC constituents not removed by centrifugation and filtration, saline was incubated with PNC. After the PNC was removed, infusion of the saline induced no significant hemodynamic changes, although profound granulocytopenia occurred. We conclude that an intact complement system, but not normal numbers of circulating granulocytes, is essential to PNC-induced hemodynamic changes. PNC generates a plasma factor in vitro that does not require an intact complement system or cellular machinery to induce granulocytopenia without hemodynamic effect.
Natural immunity to Sindbis virus is influenced by host tissue sialic acid content.
Recent studies have shown that the sialic acid content of Sindbis virus influences both its ability to active the alternative pathway in vitro and its susceptibility to complement dependent clearance from the bloodstream in vivo. Other studies have shown that the sialic acid content of Sindbis virus is determined by the host in which it is propagated. Because individuals vary in their cell surface sialic acid content, it is possible they also vary in their ability to defend themselves against Sindbis virus infection by virtue of their ability to modify the virus sialic acid content and thereby the capacity of the virus to activate the alternative pathway. To test this hypothesis, outbred Swiss mice were injected subcutaneously with Sindbis virus. There was a significant positive correlation between the level of viremia 18 hr after infection and the sialic acid content of the host's erythrocytes. In addition, animals with erythrocyte sialic acid levels equal to or greater than the mean had a higher level of viremia than animals with erythrocyte sialic acid levels less than the mean. Finally, animals that had muscle sialic acid levels equal to or greater than the mean had a higher incidence of viremia than animals with muscle sialic acid levels less than the mean. These studies suggest that the amount of tissue sialic acid in an individual host influences its ability to resist Sindbis virus infection.
Surface determinants of Haemophilus influenzae pathogenicity: comparative virulence of capsular transformants in normal and complement-depleted rats.
In an assessment of the contribution of different capsular polysaccharides to the pathogenicity of Haemophilus influenzae, the virulence of H influenzae strain Rd and of a series of capsular transformants (types b, c, d, and f) of strain Rd was compared and in normal and complement-depleted rats. After intraperitoneal inoculation, the encapsulated transformants were strikingly more virulent than strain Rd, although their pathogenicity varied; type b was the most virulent, type c the next most virulent, type f less virulent, and type d the least virulent. C3 depletion enhanced the susceptibility of rats to systemic infection but did not influence the relative virulence of the transformants. Comparative studies of clearance showed efficient intravascular survival of type b, c, and f transformants but not of the type d transformant or strain Rd; C3 depletion enhanced the intravascular survival of type d. Further analysis of the capsular transformants revealed differences in the electrophoretic characteristics of their lipopolysaccharides. These studies indicate that elaboration of a unique capsular polysaccharide may not be a sufficient explanation for the greater virulence of H influenzae type b and that lipopolysaccharide may contribute to the mediation of the differential pathogenicity of the various serotypes.
Meningococcemia in genetically determined deficiency of the second component of complement.
Explore the source record for details and available documents.
Familial systemic lupus erythematosus: immunogenetic studies in eight families.
Familial SLE provides a unique opportunity to study the relationships of previously associated genetic factors (HLA and complement component deficiencies) to the occurrence of SLE, other immune disorders, and autoantibodies in families. Thus, eight families containing two or more affected members with SLE (n = 22) and their relatives (n = 40) were examined for HLA genotypes, complement components and autoantibodies. Among the 40 non-SLE relatives, 7 (18%) had other immune diseases, including thyroid disease in 2, rheumatoid arthritis in 2, ITP in 2, and Henoch-Schönlein purpura in one. This compared to 4 of the 22 SLE patients (also 18%) of whom 3 had thyroid disease and one PSS. Eleven non-SLE relatives (28%) had ANA, which was in high-titer in 5 (13%). Eleven (28%) had antibodies to ssDNA, and one had a BFP. Only the SLE patients had antidsDNA, Ro(SSA), Sm or nRNP. A heterozygous C2 deficiency (C2D) was found in only one of the seven kindreds studied, and followed an A25, B18 DR7 haplotype. Heterozygous C2D, however, was inherited by only one of three family members with SLE. HLA-DR2 occurred in 36% and DR3 in 36% of SLE patients, which was not significantly different from either non-SLE family members or unrelated local controls. Sib pair analysis of seven sets presented here and seven from two published reports, demonstrated only random distribution with SLE. Similarly, other immune disorders and autoantibodies followed no consistent HLA haplotypes or DR specificities. Interestingly, DR2 and/or DR3 were found in five of the six patients (83%) having anti-ro(SSA) antibodies (p = NS). These data strongly suggest that genetic factors (other than HLA and complement component deficiencies) and/or environmental factors are necessary for the expression of SLE and other immune abnormalities in lupus families.
Participation of complement in host defense against capsule-deficient Haemophilus influenzae.
To investigate the role of complement in immunity to capsule-deficient Haemophilus influenzae, rats were depleted of C3 with cobra venom factor and challenged with three different strains of capsule-deficient H. influenzae. Two of them (Rd and U1) did not elaborate type b capsular antigen, whereas the other (S2) elaborated 0.16% of the amount made by its type b parent strain. Depletion of C3 significantly enhanced early intravascular bacterial survival after intravenous inoculation and strikingly increased the susceptibility of rats to infection with capsule-deficient H. influenzae. After intraperitoneal inoculation with strain Rd or U1, C3-depleted rats developed bacteremia, whereas control rats did not; challenge with strain S2 resulted in transient bacteremia in normal rats and in death in C3-depleted animals. To determine whether the greater virulence of strain S2, as compared with strain Rd or U1, was accounted for by the small amounts of capsular antigen it elaborated, we also compared its relative virulence to that of three genetically closely related capsule-deficient variants elaborating either small amounts of type b capsule or producing no detectable b antigen. No difference in virulence was observed among these four variants; all C3-depleted rats inoculated developed bacteremia of similar magnitude followed by similar mortality rates. These studies demonstrate a significant role for complement in host defense mechanisms against capsule-deficient H. influenzae and suggest that interstrain differences of virulence are not attributable to residual elaboration of small amounts of type b capsule.
Extensive verrucosis in primary immunodeficiency diseases.
Explore the source record for details and available documents.
Prolonged excretion of group A coxsackievirus in an infant with agammaglobulinemia.
Explore the source record for details and available documents.
Participation of complement in host defense against encapsulated Haemophilus influenzae types a, c, and d.
Each of the serotypes of Haemophilus influenzae (types a to f) may colonize the respiratory tract of humans, but only type b strains commonly cause invasive systemic infections. To investigate the role of complement in immunity to encapsulated non-type b strains, rats were depleted of C3 with cobra venom factor and challenged with representative serotypes of H. influenzae (type a, b, c, or d) by different routes. After intravenous challenge, rats depleted of C3 had a greater incidence and magnitude of bacteremia with each of the serotypes when compared with normal controls. Intraperitoneal inoculation of type b organisms resulted in meningitis in normal and C3-depleted rats, but only C3-depleted, and not normal, rats developed meningitis after inoculation of serotype a, c, or d. In contrast to systemic inoculation, intranasal challenge with the different serotypes resulted in bloodstream invasion and meningitis only after challenge with type b organisms. These data suggest that complement plays a significant role in immunity to encapsulated, non-type b H. influenzae through its effect on bloodstream clearance.