Study of antibodies to heterologous connective tissue, isolated from sera of patients with rheumatism]

Antibodies against connective tissue elements of various bovine organs were isolated from the sera of rheumatic fever patients with the aid of immunosorbents (bovine connective tissue extract and erythrocyte stroma). The antibody preparations obtained were not identical and contained antibodies against different antigens of bovine connective tissue. The antibody preparations failed to react with human connective tissue components.     

Use of immunoenzyme analysis for evaluating the specificity of the antibody recognition of the antigenic determinants of Streptococcus group A

Investigations carried out with the use of a CELIA system have revealed that antibodies in the sera of patients with primary erysipelas and antibodies in rabbit antiserum to the ribosomes of group A streptococcus specifically bind with adsorbed streptococcal ribosomes, recognizing the antigenic determinants of streptococcal ribosomes, which differ from those of individual Gram-negative prokaryotes (Escherichia coli, Shigella sonnei, Shigella flexneri, Salmonella minnesota). The modified CELIA system used in this investigation has made it possible to find out that antibodies in the sera of patients with primary erysipelas and antibodies in rabbit ribosomal antiserum bind with different antigenic determinants of the ribosomes of group A streptococcus.     

The immune response to group B streptococcus type III capsular polysaccharide is directed to the -Glc-GlcNAc-Gal- backbone epitope

The structures of the branched capsular polysaccharides of group B streptococcus type III (GBSIIIPS) and Streptococcus pneumoniae type 14 (Pn14PS) are identical apart from the (α2→3)-linked sialic acid in the side chains of GBSIIIPS. The present study tries to determine the minimal epitope in GBSIIIPS, using both a panel of anti-Pn14PS mouse sera and sera of humans vaccinated with either Pn14PS or GBSIIIPS. Type-specific Pn14PS antibodies that recognize the branched structure of Pn14PS have a low affinity for the native GBSIIIPS. Desialylation of GBSIIIPS results in dramatically higher affinity of anti-Pn14PS antibodies. Epitope specific anti-Pn14PS mouse antibodies and human sera of PCV7 vaccinees only recognized structures with the branching element -Glc-(Gal-)GlcNAc-, in particular -Gal-Glc-(Gal-)GlcNAc- in Pn14PS. On the other hand anti-GBSIIIPS human antibodies recognize predominantly the linear structure in the backbone of Pn14PS or GBSIIIPS, i.e., -Glc-GlcNAc-Gal-. This difference in antigenicity of Pn14PS and GBSIIIPS is in agreement with the difference in flexibility of the two polysaccharides caused by the presence or absence of sialic acid.     

Determination of antibodies to Streptococcus group A polysaccharide by an immunodiffusion method in human sera in various pathological processes of streptococcal etiology

A method for the assay of antibodies to the specific antigenic determinant of group A streptococcal polysaccharide (A-polysaccharide) in human sera was developed. The sera were tested in the precipitation test in agar gel with different doses of A-polysaccharide. The presence of a high level of the above-mentioned antibodies is indicative of infection caused by group A streptococcus, but not streptococci of other groups or by the L-forms of streptococci. In 87.5% of patients with primary rheumatism a high level of antibodies to the specific antigenic determinant of A-polysaccharide was detected during the first day of the disease, which confirms most convincingly the etiological role of group A streptococcus in rheumatism. Considerable differences in the level of antibodies to A-polysaccharide in the active and non-active phases of rheumatism have been established, which makes it possible to use the presence of a high level of these antibodies as an indicator of the rheumatic process activity. A considerable percentage of sera with a high level of antibodies to A-polysaccharide was also detected in erysipelas and acute glomerulonephritis patients.     

Immunochemical analysis in research on antibodies to Streptococcus group A ribosomes

The use of the competitive enzyme immunoassay (EIA) has made it possible to demonstrate that antiserum to the ribosomes of group A streptococcus, type 29 M, contains antibodies to homologous protein M and does not contain antibodies to group A polysaccharide, lipoteichoic acid and peptidoglycan. Ribosomal antiserum has been found to bind with the surface of heterologous type M streptococci in EIA, while forming no precipitation lines with heterologous proteins M in the double immunodiffusion test, which indicates that the binding of ribosomal antibodies with the surface of group A streptococcal cells is not type-specific. Antibodies to the ribosomes of group A streptococcus recognize some of the antigenic determinants of E. coli ribosomes.     

Reactions of antibodies to streptococcus group A antigens with fibroblasts of human heart interstitial connective tissue]

The indirect immunofluorescence method was applied to the study of the serum of rabbits immunized with fractions containing nontype-specific antigens of streptococcus, group A, belonging to the cell wall proteins. Antibodies reacting with fibroblasts of the interstitial connective tissue of the human heart were revealed in the sera. On the basis of the experimental results of absorption a supposition was put forward on the presence of a cross-reacting antigen common with the fibroblast antigen in some hydrochloric extracts obtained from streptococci, groups A and C.     

Dynamics of formation of antibodies of different immunochemical types to several L-forms and mycoplasmae]

Both IgM- and IgG-immunoglobulins were revealed in the sera of rabbits immunized with L-forms of hemolytic streptococcus, group A, and Mycoplasma fermentans. In the process of immune response the hemagglutinin titres proved to increase, with IgG antibodies prevalence in the hyperimmune sera.     

Haemagglutination-inhibiting and neutralizing antibodies to type 3 and 5 rhinoviruses in human sera and nasal washings.

Presence of antibodies to RV 3 and RV 5 was tested by HIT and NT in 60 human sera. Antibodies to RV 3 were detected in 23 sera by HIT in a titre range of 1:4--1:64 and in 19 sera by NT in a titre range of 1:4--1:256. Antibodies to RV 5 were detected in 31 sera by HIT in titres of 1:4--1:268 and 27 sera by NT in the same titre range. In a group of 22 persons with unequivocal serum antibodies nasal secretory antibodies were found in 11 subjects in titres of 1:4--1:32. In a group of 16 persons without detectable serum antibodies, presence of secretory antibodies (titre 1:4) was only found in four cases.     

Immunohistochemical study of cardiolipin and phosphatidylinositol in mouse liver]

Localization of two phospholipid haptens--cardiolipin and phosphatidylinositol--in mouse liver sections was studied by the indirect method of fluorescent antibodies. Two types of liver sections--paraffin and cryostat, and two type of fixation--in acetone, and in the acetone, buffer, and formalin mixture--were used. Antiphospholipid sera stain specifically the plasma membrane of hepatocytes and predominantly the membrane region overlooking the blood capillary. A possibility of detecting the specific phospholipid haptens depends on the method of obtaining the sections and their fixation. Two types of immunization give two types of antiphospholipid sera which differ by the stability, by the possibility of monospecific antibodies isolation from them on lipid immunosorbents, and by the types of liver section staining.     

Determination of antibodies against ribosomes and various cell wall components and detection of circulating antigens of group A Streptococcus in patients with erysipelas

The level of antibodies to the ribosomes, polysaccharide A and peptidoglycan of group A streptococcus in the blood of patients with primary, secondary, and often relapsing erysipelas was studied by means of the enzyme immunoassay with the use of the sandwich techniques. For control, the sera of healthy donors were used. In the sera obtained from all groups of erysipelas patients a significant rise in the levels of antibodies to ribosomes and peptidoglycan in comparison with the controls was revealed. An increase in the level of antibodies to polysaccharide A was revealed only in patients with frequently relapsing and secondary erysipelas. Depending on the clinical form and the duration of the disease, polysaccharide A was detected in 32-51.9% of erysipelas patients and protein-ribosomal antigen was detected in 28.6-51.9% of such patients.     

Studies of immunological activity of Aspergillus flavus Link. III. Presence of antibodies against fungal antigens in the sera of persons from selected groups

The aim of this study was to determine the occurrence of antibodies against antigens of A. flavus (APP, AEM, AS, API), A. fumigatus and A. candidus. One hundred and fifty two sera of individuals connected with industrial environment were tested, in which A. flavus was permanently isolated: 339 sera of healthy controls-blood donors of city of Poznań, and 24 sera of patients with confirmed or suspected Aspergillosis were also included in the study. The sera were tested for a presence of specific antibodies by immunoprecipitation in 1% agar gel, by using inactivated sera and above mentioned antigens. In a group of people having permanent contact with A. flavus, antibodies to antigens derived from this genus were present in 4.6% of individuals while against A. fumigatus antigens in 0% and A. candidus 0.7%. In blood donors group 5 times lower percentage of sera having anti-A. flavus antibodies was found and a complete lack of detectable antibodies for other two genera. The results of the studies of patient sera indicate a necessity of broadening a set of fungal antigens used for an investigation of this type of sera. Antibodies against A. flavus were found in three patients and for A. fumigatus in 7 patients. One patient had antibodies for both genera and two patients had antibodies against A. flavus lacking antibodies against A. fumigatus. The results of this study indicate that antigens of A. flavus should be included into serodiagnosis of Aspergillosis.     

Serosurvey for antibodies against Brucella abortus and Leptospira interrogans in pampas deer from Brazil

A survey for antibodies against Brucella abortus, and Leptospira interrogans was conducted on 17 pampas deer (Ozotocerus bezoarticus) from Pantanal Matogrossense (State of Mato Grosso do Sul, Brazil) and on 24 pampas deer from Parque Nacional de Emas (State of Goiás, Brazil). Antibodies against B. abortus were detected by plate agglutination, rose Bengal, and complement fixation tests; antibodies against Leptospira interrogans were detected by the microscopic agglutination test. All sera were negative for B. abortus antibodies and all deer sera from Parque Nacional de Emas were negative for L. interrogans antibodies. Four (24%) of 17 sera from Pantanal Matogrossense were positive for L. interrogans serovar (n = 2) hardjo, wolffi (n = 1) and mini (n = 1). While these diseases do not appear to be of major importance to the health status of Pampas deer, it appears that deer are reservoir for leptospirosis in one of the study areas.     

Human antibody responses to Brugia malayi antigens in brugian filariasis.

Human antibody responses to Brugia malayi antigens were studied with sera from a Brugia endemic area in South India. Patients with clinical filariasis had significantly higher IgE and lower IgG4 levels to adult worm antigens than people with asymptomatic microfilaraemia. Intermediate antibody levels were observed in endemic normals. A majority of sera from each clinical group contained IgG antibodies to surface antigens of infective larvae (L3) by IFAT. IgG immunoblot studies did not reveal group differences in L3 antigen recognition. IgE antibodies bound to a subset of antigens bound by IgG. IgE antibodies in sera from clinical filariasis patients preferentially bound to L3 antigens at 200, 97, 68 and 58 kDa compared with sera from microfilaria carriers. These results are consistent with prior studies of antibody responses in filariasis and add new information on the targets of IgG and IgE antibodies to L3 antigens in brugian filariasis.     

Clinical immunosorbents on a basis of polymeric matrices

Immobilization of anti-IgE on space-network polymers containing aliphatic amino- and hydrazido groups as a way of producing clinical immunosorbents has been studied. Influence of active group concentration on the specific activity of the immobilized antibodies and sorption dynamics of IgE from plasma of patents are investigated. Immunosorbents can be sterilized by gamma-irradiation without any loss of capacity. It is shown that the immunosorbents can be reused after regeneration. Basing on the results obtained, acrylonitrile- divinylbenzene copolymer with hydrazido groups is considered as the most perspective for production of clinical immunosorbents.     

Absorption of Group A Streptococcus Anti-M Typing Sera with Broken Cells

Group A streptococcus anti-M typing sera that cannot be made specific by absorption with whole streptococcus cells have been absorbed with the soluble and insoluble fractions of ruptured heterologous cells. The technique has been used successfully for preparing specific anti-M sera against eight serotypes of group A streptococcus. The method involves breakage of the absorbing cells in the presence of the antiserum, and incubation of the mixture at 37 C for 1 hr, followed by 3 to 5 days of incubation at 4 C. The procedure is useful for preparing specific antiserum from certain lots of unabsorbed antiserum that otherwise would have to be discarded because of undesirable cross-reactivity.     

Determination of antibody affinity by using antigenic and antibody immunosorbents

To determine the affinity of the active centers of antibodies, cellulose immunosorbents for antibodies and antigens have been used. The fixation of serum proteins on the sorbent, the interaction of fixed antibodies with a monovalent antigen and the graphic analysis of the results thus obtained allows one to assess not only the concentration of the effective active centers on the sorbent, but also all known characteristics of antibody affinity: the average association constant K0, the common association constant Kt, the geometric association constant Kg, the average association constants which determine the affinity of different antibody groups. The use of antigenic immunosorbent permits one to determine the value of the average internal association constant K0. The determination of antibody affinity in hyperimmune antiplague sera by means of immunosorbents and red blood cells coated with capsular antigen has resulted in obtaining similar values of affinity indices.     

Antibodies to the rhamnose determinants of the polysaccharide of streptococci group A in the sera of rheumatism patients and donors]

In the sera of patients with recurrent rheumocarditis, and especially in cases of primary rheumatism, the level of antibodies to group A streptococcal polysaccharide (A-PS) has been found, according to the results of the enzyme immunoassay, to be considerably higher than in the sera of healthy donors. The level of antibodies to rhamnose determinants (RD) of A-PS has been determined by the inhibition of the immunoenzyme reaction with A-PS under the influence of a variant of group A streptococcus and rhamnose disaccharides with the bonds alpha 1-2 and alpha 1-3. In patients with recurrent rheumocarditis the level of antibodies to A-PS has been shown to be considerably higher than in healthy donors having these antibodies. In acute primary rheumatism a high level of antibodies to A-PS has been detected only in a few cases, and at the same time the prevalence of antibodies to the specific RD of A-PS, bound with beta-N-acetylglucosamine, is observed. In the sera of patients with recurrent rheumocarditis and donors having a high content of antibodies to the rhamnose site of A-PS antibodies, seemingly active against at least two RD, have been detected. In acute primary rheumatism an insignificant amount of antibodies to the rhamnose site of A-PS may probably cause the autoimmune process accompanying rheumatism. This suggestion is substantiated by the previously established capacity of these antibodies for inducing the suppression of cytotoxic cell reactions to microbial antigens.     

Antibodies to Bordetella pertussis adenylate cyclase toxin in neonatal and maternal sera

Abstract To investigate the high prevalence among infants of antibodies to Bordetella pertussis adenylate cyclase toxin (ACT), cord-blood sera were examined for antibodies to ACT, filamentous hemagglutinin (FHA) and pertussis toxin (PT) using immunoblot analysis. Antibodies reactive with ACT were the most prevalent in neonatal sera. Similar reactivity of IgG with ACT was found in each sample of a given neonatal-maternal pair, yet IgM reactive with ACT was virtually absent in neonatal sera, suggesting that antibodies to ACT are maternally derived. Antibodies to ACT might come from infection or childhood vaccination of the mothers since pertussis vaccines from all US manufacturers elicited antibodies to ACT in mice. Alternatively, these antibodies may have been elicited by a cross-reactive antigen such as Escherichia coli α-hemolysin, since all of the neonatal and maternal sera contained antibodies reactive with α-hemolysin.     

Competitive inhibition by human sera of mouse monoclonal antibody binding to glycoproteins C and D of herpes simplex virus types 1 and 2.

A competitive enzyme-linked immunosorbent assay was used to test for human antibodies to antigenic sites on herpes simplex virus (HSV) glycoproteins C and D, which are recognized by mouse monoclonal antibodies. Antibodies capable of blocking the monoclonal antibodies were detected in the human sera, and the inhibition of binding correlated with the histories of herpetic infections. The binding of monoclonal antibody to glycoprotein C of HSV type 2 was inhibited primarily by sera from patients with recurrent herpes genitalis; however, the binding of the monoclonal antibodies to gC of HSV type 1 was inhibited by sera from patients previously infected with either HSV type 1 or HSV type 2. The observations suggest that the antigenic sites defined by the mouse monoclonal antibodies are recognized by the human host.     

Measurement of Neutralizing Serum Antibodies of Patients Vaccinated with Human Papillomavirus L1 or L2-Based Immunogens Using Furin-Cleaved HPV Pseudovirions

Antibodies specific for neutralizing epitopes in either Human papillomavirus (HPV) capsid protein L1 or L2 can mediate protection from viral challenge and thus their accurate and sensitive measurement at high throughput is likely informative for monitoring response to prophylactic vaccination. Here we compare measurement of L1 and L2-specific neutralizing antibodies in human sera using the standard Pseudovirion-Based Neutralization Assay (L1-PBNA) with the newer Furin-Cleaved Pseudovirion-Based Neutralization Assay (FC-PBNA), a modification of the L1-PBNA intended to improve sensitivity towards L2-specific neutralizing antibodies without compromising assay of L1-specific responses. For detection of L1-specific neutralizing antibodies in human sera, the FC- PBNA and L1-PBNA assays showed similar sensitivity and a high level of correlation using WHO standard sera (n = 2), and sera from patients vaccinated with Gardasil (n = 30) or an experimental human papillomavirus type 16 (HPV16) L1 VLP vaccine (n = 70). The detection of L1-specific cross-neutralizing antibodies in these sera using pseudovirions of types phylogenetically-related to those targeted by the L1 virus-like particle (VLP) vaccines was also consistent between the two assays. However, for sera from patients (n = 17) vaccinated with an L2-based immunogen (TA-CIN), the FC-PBNA was more sensitive than the L1-PBNA in detecting L2-specific neutralizing antibodies. Further, the neutralizing antibody titers measured with the FC-PBNA correlated with those determined with the L2-PBNA, another modification of the L1-PBNA that spacio-temporally separates primary and secondary receptor engagement, as well as the protective titers measured using passive transfer studies in the murine genital-challenge model. In sum, the FC-PBNA provided sensitive measurement for both L1 VLP and L2-specific neutralizing antibody in human sera. Vaccination with TA-CIN elicits weak cross-protective antibody in a subset of patients, suggesting the need for an adjuvant.