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Cross-reactions between mycobacteria. II. Crossed immunoelectrophoretic analysis of soluble antigens of BCG and comparison with other mycobacteria.

Cross-reactions between Mycobacterium bovis BCG and various other mycobacteria, Nocardia asteroides, Corynebacterium pyogenes and Listeria monocytogenes were studied by incorporating antibodies against these bacteria in the intermediate gel of a crossed immunoelectrophoretic system with BCG antigen and anti-BCG antibodies. In the BCG reference system forty-four distinct antigenic components were recorded, of which thiryt-three cross-reacted with Mycobacterium tuberculosis, twenty-five with M. avium, twenty-one with M. suvalii, eighteen with M. smegmatis, Fifteen with M. nonchromogenicum, twelve with M. phlei, eight with N. anteroides and two with C. pyogenes, whereas no cross-reaction was detected with L. monocytogenes. The value of the method for characterization of mycobacterial antigens is discussed. A taxonomic system based on this method appears particularly valuable for studies of non-cultivativable mycobacteria such as M. leprae. A majority of twenty-one patients with lepromatous leprosy had anti-BCG antibodies of restricted specificity, affecting only four or five BCG antigens, although one patient had twelve anti-BCG specificities. Most of these antibodies reacted with those BCG antigens that cross-react extensively with other mycobacteria.

Antibodies, Bacterial

[Study of serologic cross reactions between the antigens of Paracoccidioides brasiliensis and Histoplasma capsulatum].

A comparative study of serologic cross-reactions between histoplasmosis and paracoccidioidomycosis was carried out using complement fixation (CF) and agar gel immunodiffusion (ID) tests. Sera from 77 patients with paracoccidioidomycosis, 38 with histoplasmosis and 50 healthy individuals were studied. The Pan American Health Organization microtiter CF test and the Hupper ID technique were employed. The antigens were prepared by the method of Ajello as modified by Negroni. Paracoccidioidin CF test was positive in all patients with paracoccidioidomycosis; 29 of these (37%) presented serologic cross-reaction with histoplasmin. Histoplasmin CF test was positive in all but one of the patients with histoplasmosis and only 4 of them (10%) showed cross-reaction with paracoccidioidin. Positive reactions with the ID test were registered in 74/77 cases of paracoccidiodomycosis, with only one case of cross-reaction (1,3%). The result for the ID test was positive in all but one of the cases of histoplasmosis, with only one cross-reaction with paracoccidiodin (2,7%). The ID test seemed to be much more specific although it could be less sensitive. It was also easier to perform and less expensive than the CF test. Nevertheless the authors urge that the CF test also be carried out whenever posible, since the information obtained is likely to be of more value in follow-up and control of patients.

Antigens, Fungal

[The background of the serological cross-reaction between Yersinia and Brucella and the possibility of differential diagnostics by the ELISA technique and the electroimmuno assay (rocket electrophoresis) (author's transl)].

A strong serological cross-reaction appears between different species of genus Brucella and Yersinia enterocolitica (Y.e.) serotype 0--9, which seriously complicates the diagnostic works of brucellosis and yersiniosis both in humans and animals. This cross-reaction makes it impossible to perform a differential serological diagnosis using common routine procedures such as the agglutination test and the complement fixation test. By the use of immunological, immunochemical and structural chemical methods it has been shown that the common antigenic structures of Brucella and Yersinia are situated in the lipopolysaccharides (LPS) of the cellwalls. In order to do structure analysis gaschromatography in combination with mass spectrometry of LPS from Brucella and Yersinia bacteria has been achieved. The results show that two monosaccharides (glucose and galactose) constitute common components of the LPS-molecules. A discovered cross-reaction between Y.e. 0--9 and Salmonella urbana (0--30) confirms the hypothesis that glucose and galactose are responsible for the serological cross-reaction between Brucella and Y.e. 0--9. An enzyme-linked immunosorbent assay (ELISA) has been developed in order to be used as a differential routine test. The results show that by using ELISA, a differentiation between antibodies against Y.e. 0--9 and B. abortus can be done with high sensitivity and accuracy. Another differential diagnostic system has been developed by the use of electroimmuno assay (rocket electrophoresis). This method raises the possibility of a rapid and a simple qualitative differentiation of antibodies against Brucella and Yersinia.

Agglutination Tests

Cross-reactions between Neisseria meningitidis and twenty-seven other bacterial species.

Cross-reactions between antigens from N. meningitidis and antigens from 27 other bacterial species from clinical isolates representing 20 different genera were studied by quantitative immunoelectrophoretic methods. A water-soluble N. meningitidis antigen preparation and a corresponding pooled rabbit antiserum regularly presenting 48 immunoprecipitates were used as reference system. Antigens from 16 other bacterial genera cross-reacted with one to four N. meningitidis antigens, whereas antigens from six other Neisseria species cross-reacted with 37 to 48 N. meningitidis antigens. Antigens from only three bacterial genera (Streptococcus, Staphylococcus and Bacteroides) did not show any cross-reactions. The degree of cross-reactivity between the antigens were 25--100%, as judged by absorption of antibodies. Three of the cross-reacting antigens reacted with normally occurring precipitating antibodies in serum from humans and rabbits.

Antigens, Bacterial

Cross reaction between bovine enterovirus and South African Territories I5 foot-and-mouth disease virus.

A bovine enterovirus (E76T) isolated from a 2-year-old bull produced serologic cross reactions to South African Territories (SAT) I5 foot-and-mouth disease virus when inoculated into guinea pigs and cattle. Cross-reacting serum titers to SAT I5 virus of 1:320 by the plaque-reduction neutralization test and 1:20 by the radial immunodiffusion test occurred in 2 steers after they were inoculated with the E76T virus. In 1 steer, maximal cross-reacting titers appeared related to a 2nd exposure to the viruses or to a hyperimmune state. Ultracentrifugation and 2-mercaptoethanol studies indicated that the cross reactions were due to immunoglobulin M antibody. Sera from guinea pigs immunized with the E76T or the SAT I5 virus cross reacted with the heterologous virus by postinoculation day 7. Cross-reacting titers had decreased markedly by postinoculation day 35, whereas the homologous virus titer remained constant. Cross reactivity of the E76T virus was primarily with the SAT I5 virus, and to a lesser degree with SAT II3. Cross reactions did not occur with representatives of the 5 other antigenic types of foot-and-mouth disease virus.

Animals

Immunochemical cross-reactions between type III group B Streptococcus and type 14 Streptococcus pneumoniae.

Serological cross-reactions between certain streptococci and some serotypes of Streptococcus pneumoniae have been reported. These studies detail the serological cross-reactivity observed between hot HCl-extracted group b streptococcus type III (GBS III) antigens and S. pneumoniae type 14 (Pn 14) polysaccharide. Similar electrophoretic migration patterns of GBS III and Pn 14 were observed when either type-specific BGS III antisera or pneumococcal omniserum was utilized to precipitate these antigens. Both the GBS III antigen and the Pn 14 polysaccharide migrated toward the cathode, whereas all other pneumococcal polysaccharides migrated toward the anode. No cross-reactions were observed between GBS III antisera and the 11 other types of pneumococcal polysaccharides. Lines of identity were observed between type-specific GBS III antisera and monospecific Pn 14 antiserum with either GBS III antigens or purified Pn 14 polysaccharide. The cross-reacting antigens of GBS III and Pn 14 appear to be identical by immunodiffusion and immunoelectrophoresis.

Antigens, Bacterial

Serological cross-reactions of leptospiral lipopolysaccharide (F4) antigen.

The serological specificity of leptospiral lipopolysaccharide (F4) antigen was examined by the technique of passive haemagglutination. F4 extracted from leptospiral serovars representative of several different serogroups showed wide cross reaction between serovars, including numerous one-way (non-reciprocal) reactions. The pattern of cross reaction was different to that of the standard leptospiral classification scheme.

Antigens, Bacterial

Cross reactions in occupational contact dermatitis. I. Aromatic amines.

Cross reactions are described between particular aromatic amines, between phenylenediamine ortho-, meta- and para-isomers and between aromatic amines and related substances. Moreover, the sensitization pattern and some sources of primary sensitivity to aromatic amines are discussed.

Allergens

Serological cross-reactions between Acinetobacter calcoaceticus and chlamydiae.

A cross-reaction between Acinetobacter calcoaceticus and chlamydiae is described. A water-soluble, heat stable, non-dialyzable antigen was extracted from Acinetobacter species by boiling. This antigen fixed complement in the presence of homologous hyperimmune sera from rabbits or guinea pigs and in the presence of heterologous human or hyperimmunized animal sera containing chlamydial antibodies. Hyperimmune antisera to the extracted antigen, or to suspensions of live acinetobacters, also reacted in complement fixation with a group-specific antigen.

Acinetobacter

Cross-reactions of normal bovine serums to foot-and-mouth disease virus in plaque-reduction neutralization and radial immunodiffusion.

Serums from 150 cattle with no known exposure to foot-and-mouth disease (FMD) virus were tested by both the plaque-reduction neutralization (PRN) technique and the radial immunodiffusion (RID) technique to evaluate the significance and the extent of cross-reactions in these tests. Serums from 30 cattle from each of 5 locations were tested against representative viruses of each of the 7 types of FMD virus. High levels of cross-reactions with both the RID and PRN techniques were found in serums of specific groups. Higher levels of cross-reactions were noted for Asia, SAT I, SAT II, and SAT III than for A, O, or C viruses with the PRN technique, and higher levels for SAT II and SAT III with the RID technique. The reactions for most serums appeared to be quite specific; a given serum was often reactive with only 1 or 2 viruses.

Animals

Nature of cross-reaction between hCG and anti-oLH serum and development of a radioimmunoassay to measure hLH specifically in the presence of hCG.

Immunological cross-reaction between hCG and anti-oLH sera has been demonstrated using radioimmunoassay techniques. The results indicate that this cross-reaction is incomplete and that the anti-oLH sera used have the ability to distinguish between LH and hCG. Following absorption with purified hCG, anti-oLH serum was used to develop a heterologous radioimmunoassay "[125I]iodo-hLH + anti-oLH serum" (H-O, RIA) which specifically and selectively measures hLH in serum samples containing both hLH and hCG. In this radioimmunoassay hCG and subunits of hCG do not cross-react with hLH, in the range in which these hormones are present in human serum under physiological conditions. Other hormones such as hPL, hPRL, hGH, hFSH, hTSH, and GnRH do not interfere with the measurement of LH by radioimmunoassay. The sensitivity of the assay was 1.5 mIU (25 ng) per ml (LER 907 standard), and the inter- and intra-assay coefficients of variations for samples were 10.83% and 8.4%, respectively. The recoveries of hLH added to pregnancy serum containing an hCG concentration of 8.55 IU/ml were in the range 95-108%. Determination of LH content of human pituitary extracts by H-O RIA gave values which were in close agreement with those derived by bioassay (indices of discrimination 0.72-1.12). Serum LH patterns in women during normal menstrual cycles as well as in amenorrheic patients who received GnRH treatment are comparable to those reported by other investigators using other radioimmunoassay systems. Serum samples obtained during the first trimester of pregnancy, when analyzed by H-O RIA, showed basal LH levels.

Amenorrhea

Occurrence of cross reactions to foot-and-mouth disease virus in normal swine sera.

Sera from 101 swine never exposed to foot-and-mouth disease virus were tested by the plaque-reduction neutralization (PRN) and radial immunodiffusion techniques for cross-reactions to 5 types of foot-and-mouth disease viruses. Depending on the group of sera and the virus used, the percentage of sera cross-reacting at low levels varied from 0 to 50% with the PRN technique and 0 to 20% with the radial immunodiffusion technique. 5erum-neutralization tests in mice support the finding of neutralizing antibody by the PRN technique. Ultracentrifugation and 2-mercaptoethanol studies indicate that the cross-reactions are the result of immunoglobulin M or similar macroglobulins.

Animals

Streptococcal M protein extracted by nonionic detergent. III. Correlation between immunological cross-reactions and structural similarities with implications for antiphagocytosis.

Three immunologically cross-reactive and non-cross-reactive streptococcal M proteins were analyzed by a chromatographic tryptic peptide mapping system. The results indicate that cross-reactions correlate with the extent of structural similarity among the M protein molecules analyzed. The data also reveal that free lysine is released by the action of trypsin from these three M proteins, suggesting a common lys-lys or arg-lys sequence. In addition, only one peptide has been found to be common within all three M types. This limited structural relatedness among the three M proteins examined indicates that sequence variation plays a major role in the immunological specificity of the M antigens. However, despite sequence variation, all M protein molecules have a common antiphagocytic activity. The fact that no common opsonic antibody has yet been found, even against limited M types, argues against this biological activity being solely the result of a common sequence. Based on these data, it is suggested that the antiphagocytic effect of M protein may be due to a conformationally created environment on the surface of the molecule which is selected by both immunological and biological pressure.

Antigens, Bacterial

Immunochemistry of serum albumin. VI. A dynamic approach to the immunochemical cross reactions of proteins using serum albumins from various species as models.

Antisera against bovine serum albumin were raised in two rabbits. Serial bleedings were obtained at different times after the first immunization, and antisera from these serial bleedings were not mixed but were kept and studied separately. The immunochemical cross-reactions of these antisera with serum albumins from bovine, goat, sheep, porcine, horse, human and chicken were determined by immunoadsorbent studies. These were done by titration so that the values of maximum (plateau) binding by each albumin of radioiodinated antibodies were determined. In each rabbit, the immunochemical cross-reactivity was not static but increased progressievly with time after the first immunization. In the interval 7 days to 398 days the increases in cross-reaction were extremely large. pH dissociation studies revealed that, together with the increase in cross-reactivity of a given albumin with time after immunization, there was a restriction in the antibody heterogeneity towards populations possessing higher affinity. These results provide a rational explanation for the different values of cross-reactivities for a given albumin from different laboratories. The findings are analyzed in relation to the antigenic structure of albumin and their significance in evolutionary studies discussed.

Animals

Cross-reactions between Bordetella pertussis and twenty-eight other bacterial species.

Cross-reactions between B. pertussis and 28 other bacterial species were studied by various quantitative immunoelectrophoretic methods. A sonicated B. pertussis antigen preparation and a corresponding pooled rabbit antiserum were used as reference system. Two of the B. pertussis antigens were cross-reactive with antigens from 17, respectively 19, other bacterial species mainly gram-negative species. As judged by absorption of antibodies, the degree of cross-reactivity of these B. pertussis antigens with antigens from other species was found to be in the range 25-30%. Antigens from B. parapertussis and B. bronchiseptica were found to cross-react very extensively with B. pertussis, and only 4, respectively 2, of the 44 antigens of the B. pertussis reference system could not be absorbed with antigens from these two Bordetella species.

Antibodies, Bacterial

Cross reactions of normal bovine sera with foot-and-mouth disease virus: incidence, duration, and effect of shipping stress.

Serum samples were obtained from 30 Hereford steers in an area known to be free of foot-and-mouth disease (FMD) viruses as follows: before shipment and 4 times during a 70-day period after shipment; the sera were tested for the presence of cross-reacting antibody to various viruses. Percentages of sera containing cross-reacting antibody to FMD virus detected by the plaque-reduction neutralization and the radial immunodiffusion techniques were higher for the FMD viruses Asia and SAT I5 than for the FMD viruses A5, O1, and C1. Cross-reacting antibody was usually of low titer and usually present in only 1 or 2 consecutive serum samples. The incidence of cross reactions increased after stress of shipping and thus an infective agent may be responsible. These results were compared with results from sera collected from Herefords and Holstein-Friesians in a 2nd area; results did not indicate that Herefords have an excess of cross reactions with FMD viruses.

Animals

Cross-reactions between tryptic polypeptides of staphylococcal enterotoxins B and C.

The strong cross-reactions demonstrated for staphylococcal enterotoxins B (SEB) and C1 (SEC1) by measurement of antigen-binding capacity were reflected in well defined polypeptides obtained by limited tryptic digestion from SEB and SEC1. Two antigenic determinants on each enterotoxin were capable of reacting with heterologous antibody, one on the first 57 amino acids and one on the last 150 residues of the polypeptide backbone. The larger, carboxyl terminal polypeptides bound efficiently to homologous antiserum but about two orders of magnitude less efficiently to heterologous antibody. The amino terminal peptides showed only weak homologous binding but nearly comparable heterologous binding. It is proposed that the determinant on the amino terminal polypeptides is largely responsible for the strong reciprocal binding of the intact enterotoxins and that their low antigen-binding capacity is due to a random or a structurally distorted conformation in solution.

Animals

Use of RAST technique in wasp sting hypersensitivity. Cross-reactions between various insect antigens are specially considered.

Clinical hypersensitivity to wasp stings was found to be fairly well correlated with the presence of serum IgE-antibodies against yellow jacket venom as detected by the RAST technique. Such antibodies were never found in a control group of non-allergic blood donors, but they were detected in a surprisingly large proportion of patients with bee sting allergy without known allergic reactions to wasps. Studies using RAST inhibition technique failed to prove cross-reactions between bee and wasp venoms. Considerably better results were obtained when venom antigens instead of whole body antigens were used in the RAST. RAST inhibition studies suggested that IgE-antibodies detected with RAST using whole body antigen are directed against bee venom constituents in the whole body extract.

Antibodies