Serological analysis and pathogenic potentials of Wangiella dermatitidis isolated from bats

Methods for the preparation of antigens from clinically isolated cultures of Aspergillus were standardized. Sera from 25 suspected cases of pulmonary aspergillosis were tested against antigens prepared by us, from 4 strains of A. fumigatus and one strain of A. flavus, using the Ouchterlony double diffusion and immunoelectrophoretic techniques.Of the 25 sera tested, 18 reacted positively with antigens of A. fumigatus, one with A. flavus and 2 with both these species. Antigens of two non-pathogenic Aspergilli included in the study failed to react with any of the sera. Our antigen preparations gave more numerous as well as sharper precipitin lines than the commercial Bencard antigens which were used for comparison. Moreover, mycelial antigens from 48 to 96 h old cultures revealed precipitin lines comparable to that of the routine, 4 week old culture filtrate antigens, thus suggesting that the incubation period for obtaining antigens could be cut down considerably.Memoir No. 323 from the Centre of Advanced Study in Botany.Deceased     

T Cell Response to Borrelia garinii,Borrelia afzelii,and Borrelia japonica in Various Congenic Mouse Strains

The infectivity and T cell response to Borrelia garinii SIKA2, Borrelia afzelii BFOX, and Borrelia japonica 0612, the organisms that cause Lyme disease in Japan, were examined in various inbred and congenic strains of mice. Infectivity differed among the species: B. garinii SIKA2 and B. afzelii BFOX were each able to infect 90% to 100% of C3H/He mice; B. japonica 0612 was able to infect only 20% of C3H/He mice. The pattern of infectivity to various inbred and congenic strains of mice may influence the pathogenicity of the organism and the clinical signs of Lyme disease. Cross-reactivity between Borrelia antigens was observed, but there was no cross-reactivity between Borrelia antigens and Leptospira antigens. We evaluated the genetic control of the delayed-type hypersensitivity (DTH) reaction in the form of footpad swelling produced by Borrelia antigens using viable or sonicated bacteria as sensitization. Differences in strains of mice infected by viable antigen were observed. However, all strains of mice showed a strong DTH reaction using sonicated antigens without genetic background. A DTH reaction in the form of footpad swelling did not appear to be associated with genetic background. The footpad reaction was mediated by CD4⁺8^? and Ia^? T cells, as revealed by in vitro monoclonal antibody treatment. However, CD8⁺ T cells did not suppress footpad swelling. These results indicate that many antigenic epitopes of the Borrelia spirochete can stimulate the DTH reaction.     

Serological characteristics within the genus Desulfovibrio

Antisera were prepared against one strain each of Desulfovibrio desulfuricans, D. vulgaris and D. salexigens. The antisera were tested for cross reactivity against 36 heterologous Desulfovibrio strains by both agglutination titration and by double immunodiffusion precipitin plates.Generally no cross-reaction was demonstrated by agglutination even between heterologous strains of the same species, suggesting that the surface antigens of Desulfovibrio are highly specific. In immunodiffusion plates a single apparently genus-specific surface antigen could be shown to be present in all but two of the strains tested. Although other common precipitin bands showed the presence of some antigens common between heterologous strains these appeared to be randomly distributed among the strains tested, with the exception of one band shown to be generally specific to strains of D. salexigens. With this exception no other precipitin band could be shown to be consistently specific to any other species, nor consistently common to more than one species.     

Antigenic Analysis of Virulent and Avirulent Strains of Trichomonas gallinae by Gel Diffusion Methods*

SYNOPSIS. Antigenic constitution of 5 Trichomonas gallinae strains and substrains was analyzed by gel diffusion technics. Fresh isolates of the very virulent JB and of an avirulent SG strain as well as avirulent substrains JBC and SGC, derived from JB and SG respectively by prolonged in vitro cultivation, were used in the experiments. An originally avirulent AG strain that was attenuated still further and lost its infectivity for pigeons during many years of serial transfers in nonliving media also was analyzed. Two major groups of antigens, A and B, were differentiated on the basis of precipitin line patterns formed in gel diffusion reactions involving the 5 strains and substrains and antisera prepared in rabbits against each of these trichomonad stocks. Group A was subdivided further into subgroups [A] and (A). JB, JBC, AG, and SGC trichomonads appeared to share all or nearly all antigens of both these subgroups, but AG strain contained some unique [A] and (A) antigens in addition to those which it had in common with the remaining 4 strains and substrains. Group B antigens were divided into 5 subgroups, B₁ to B₅. The complete B₁ antigenic complex was found in JB and JBC trichomonads and part of this complex was present also in SG strain and SGC substrain. In all instances, subgroup B₁ antigens stimulated production of specific antibodies in rabbits and combined with these antibodies present in immune sera. The complete B₂ antigenic complex was found only in JBC substrain. Some subgroup B₂ antigens were present also in JB trichomonads. Very few of these, however, were capable of stimulating antibody production in rabbits. The more numerous B₂ elements of JB strain that did not stimulate immunologic responses in rabbits, might be in the form of incomplete hapten-like antigens. All subgroup B₂ antigens found in JB strain represented only a portion of the B₂ complex associated with JBC substrain. Subgroup B₂ was characteristic of SG and SGC trichomonads, the latter substrain differing from the parental SG strain in the levels of both B₂ and B₁ antigens; these differences, however, were purely quantitative. JB strain reacted with some of subgroup B₃ antibodies present in SG and SGC antisera, but failed to stimulate antibody formation against any of these antigens in rabbits. The B₃ elements of JB trichomonads might be incomplete antigens. AG strain was characterized by having B₄ and B₅ antigenic complexes. The very small part of subgroup B₄, represented by a weak precipitin line in reactions between JB strain or JBC substrain and anti-AG serum, suggested the presence of some incomplete B₄ antigens in these trichomonads. Irrespective of whether freshly isolated avirulent strains or substrains attenuated by prolonged in vitro cultivation are examined by gel diffusion, such organisms are found richer in subgroup B antigens than the fully virulent JB trichomonads. All the results suggest that there may be a direct relationship between antigenic constitution and virulence of T. gallinae strains.     

Species-specific and cross-reactive antigens of Entamoeba

Specific and cross-reactive antigens were defined in four species of Entamoeba: invadens, moshkovskii, Laredo and histolytica strains HM1, HM3, HM38 and HK9. Among these species extensive common reactivities were observed by immunoblot. Eight E. histolytica antigenic markers were revealed after blocking common specificities with antigens of other Entamoeba species. A monoclonal antibody (mAb) defined two protein markers of E. histolytica, M, 29 and 25 kDa. The four strains of E. histolytica, which varied in virulence as determined by the development of liver abscesses in hamsters, showed the same antigenic patterns with the mAb and with polyclonal antibodies.     

Group antigens in slow-growing rhizobia

Summary Internal group antigens of several slow-growing and fast-growing Rhizobium strains were tested by gel-diffusion against antisera to three strains of Rhizobium japonicum. At least one, generally two common antigens were found in 13 strains of R. japonicum, 4 strains of R. lupini, 4 strains isolated from cowpea and two slow-growing strains isolated from Lotus. Forty-six fast-growing rhizobia (including two from Lotus and 4 from Leucaena leucocephala) were clearly distinguished from the slow-growing strains in tests with the same antisera. They were wholly negative (9) or gave a much weaker non-identical line with one antiserum (24 strains), two antisera (8) or three antisera (5). The 5 strains of agrobacteria grouped with the fast-growing rhizobia.     

Linkage of a 7S RNA sequence and kappa light chain genes in the mouse

A mouse 7S RNA cDNA plasmid clone was employed to identify and map DNA restriction fragment variants using recombinant inbred (RI) and congenic mouse strains. More than a dozen such restriction variants were identified and mapped to different regions of the mouse genome. One such variant, designated Rn7s-6, showed close linkage to the Ly-2,3-Igk-V (T lymphocyte antigens 2 and 3, kappa immunoglobulin variable region) cluster of markers on chromosome 6. No recombinants were detected among three of these markers in 59 RI strains. On the basis of these data, the Rn7s-6 sequence may be placed within 1.3 centimorgans of Ly-3 and one of the Igk-V-region markers, Igk-Efl. Two mouse stocks with previously identified crossovers within the Ly2,3-Igk-V region were used to sublocalize Rn7s-6. The results are consistent with the gene order (Ly-2, Ly-3)-(Rn7s-6, Igk-Efl)-Igk-Ef2. Several mouse plasmacytomas, known to have various parts of the kappa chain complex deleted, retain the Rn7s-6 sequence. The Rn7s-6 variant is a plus/minus variant; no sequence allelic to Rn7s-6 is found in inbred strains that share the Ly-3 ^a-Igk-Efl^a haplotype.     

Antigenic diversity of cytochromes c3 from the anaerobic,sulfate-reducing bacteria,Desulfovibrio

An indirect enzyme-linked immunoadsorption assay (ELISA) was developed for cytochrome c₃ using antisera to the cytochromes fromDesulfovibrio africanus Benghazi, Desulfovibrio vulgaris Hildenborough andDesulfovibrio salexigens British Guiana. The ELISA system was used to test for cross-reactions between these antisera and the heterologous antigens. In contrast to previous experiments using the Ouchterlony technique, all of the cytochromes c₃ tested exhibited some degree of cross-reaction. Considerable variation was seen in cross-reactions for cytochromes c₃ from differing strains ofD. desulfuricans. This observation raises questions about the taxonomic relatedness of these strains. No cross-reaction was seen with eukaryotic cytochrome c or withD. vulgaris cytochrome c₅₅₃. The data demonstrate that cytochrome c₃ is capable of undergoing nonprecipitating cross-reactions, and thus may not be as immunologically unique as was once thought.Abbreviations ELISA Enzyme-linked immunoadsorption assay     

Genetic and immunological characterization of naturally occurring recombinant B3 rats

The B-stock population of rats was bred for homozygosity at the loci controlling coat color. In this process, theAg-B1 andAg-B3 haplotypes became fixed in Hardy-Weinberg equilibrium. Extensive immunization and absorption studies showed that the specificities in the B-stock rats homozygous for theAg-B1 haplotype were the same as those found in the inbred F344 strain (Ag-B1), and that the specificities in the rats homozygous for theAg-B3 haplotype were the same as those found in the inbred BN (Ag-B3) strain. A homozygous line derived from the rats carrying theAg-B3 haplotype (B3) has the mixed lymphocyte reactivity and antibody responsiveness to poly (Glu⁵²Lys³³Tyr¹⁵) characteristic of the inbred strains in theAg-B4 group. Thus, it represents a naturally occurring recombination between the loci controlling MLR and immune responsiveness, on the one hand, and those controlling the Ag-B antigens on the other. Antibody responsiveness segregated with theAg-B3 haplotype in crosses between the B3 homozygotes and the low responder BUF and M520 strains; hence, this recombination is a stable one. There was no linkage of antibody formation or haplotype to coat color. The finding of a strain with a naturally occurring recombination in the major histocompatibility complex between the loci controlling mixed lymphocyte reactivity and the Ag-B histocompatibility antigens provides evidence for the separateness of these loci. Since the portion of the genetically determined mechanism controlling antibody responsiveness which is linked to the MHC was that characteristic of the MLR type, it too must lie outside the region defined by the serological specificities of theAg-B haplotype.     

Identification of Helicobacter pylori strain cagPAI+ and cagPAI- Antigens by IgG antibodies from sera of experimentally colonized meriones unguiculatus (Mongolian gerbils)

Background: Mongolian gerbils that are experimentally infected with Helicobacter pylori develop a chronic inflammation that is similar to natural infections in humans. The aim of this study was to compare the antigens of H. pylori cagPAI+ and cagPAI? strains that are expressed during Meriones unguiculatus colonization. Materials and Methods: We identified H. pylori cagPAI+ and cagPAI? strain antigens via Western blotting of samples from Mongolian gerbils that were subjected to unique, mixed, and sequential bacterial infections. Results: The antigens from the J99/CG3 (cagPAI+) strain had a lower molecular weight than the antigens from the 251F/CG3 (cagPAI?) strain. There were fewer identified antigens in the single unique infections compared with the mixed and sequential infections. The number of recognized antigens that had a frequency of recognition >60% was higher for the simultaneous and sequential infection groups compared with the single infection group. A 57‐kDa antigen was present in >60% of the samples and four of the five experimental groups. Antigens specific to each bacterial strain were identified; the 190‐ and 158‐kDa antigens appear to be specific for cagPAI?, and the 70‐kDa antigen appears to be specific for cagPAI+. Conclusions: In this study, we identified antigens that are common and specific to the H. pylori cagPAI+ and cagPAI? strains.     

Immunological relationship of extra-cellular polysaccharide antigens produced by different mould species

Moulds are able to produce extracellular polysaccharide antigens which are heat-stable and almost genus specific. Of 44 different strains of Penicillium 41 (93%) and all 12 strains of Aspergillus tested produced detectable quantities of an immunologically related antigen. Additionally 10 of these 56 strains produced an antigen immunologically related to the antigen produced by the genera Mucor and Fusarium. Immunologically different, but genus-specific antigens were produced by each of the species belonging to the genus Geotrichum, Fusarium, Cladosporium, Mucor and Rhizopus. The antigens produced by Mucor and Rhizopus, however, were immunologically related.     

The genetic origin of minor histocompatibility antigens

The purpose of this study was to elucidate the genetic origin of minor histocompatibility (H) antigens. Toward this end common inbred mouse strains, distinct subspecies, and species of the subgenus Mus were examined for expression of various minor H antigens. These antigens were encoded by the classical minor H loci H-3 and H-4 or by newly identified minor H antigens detected as a consequence of mutation. Both minor H antigens that stimulate MHC class I-restricted cytotoxic T cells (Tc) and antigens that stimulate MHC class II-restricted helper T cells (Th) were monitored. The results suggested that strains of distinct ancestry commonly express identical or cross-reactive antigens. Moreover, a correlation between the lack of expression of minor H antigens and ancestral heritage was observed. To address whether the antigens found on unrelated strains were allelic with the sensitizing minor H antigens or a consequence of antigen cross-reactivity, classical genetic segregation analysis was carried out. Even in distinct subspecies and species, the minor H antigens always mapped to the site of the appropriate minor H locus. Together the results suggest: 1 minor H antigen sequences are evolutionarily stable in that their pace of antigenic change is slow enough to predate subspeciation and speciation; 2 the minor H antigens originated in the inbred strains as a consequence of a rare polymorphism or loss mutation carried in a founder mouse stock that caused the mouse to percieve the wild-type protein as foreign; 3 there is a remarkable lack of antigenic cross-reactivity between the defined minor H antigens and other products.     

Involvement of Mhc Loci in immune responses that are not Ir-gene-controlled

Twenty-nine randomly chosen, soluble antigens, many of them highly complex, were used to immunize mice of two strains, C3H and B10.RIII. Lymphnode cells from the immunized mice were restimulated in vitro with the priming antigens and the proliferative response of the cells was determined. Both strains were responders to 28 of 29 antigens. Eight antigens were then used to immunize 11 congenic strains carrying different H-2 haplotypes, and the T-cell proliferative responses of these strains were determined. Again, all the strains responded to seven of the eight antigens. These experiments were then repeated, but this time -antibodies specific for the A (AA) or E (EE) molecules were added to the culture to block the in vitro responsiveness. In all but one of the responses, inhibition with both A-specific and E-specific antibodies was observed. The response to one antigen (Blastoinyces) was exceptional in that some strains were nonresponders to this antigen. Furthermore, the response in the responder strains was blocked with A-specific, but not with E-specific, antibodies. The study demonstrates that responses to antigens not controlled by Irr genes nevertheless require participation of class II Mhc molecules. In contrast to Ir gene-controlled responses involving either the A- or the E-molecule controlling loci (but never both), the responses not Ir-controlled involve participation of both A- and E-controlling loci. The lack of Ir-gene control is probably the result of complexity of the responses to multiple determinants. There is thus no principal difference between responses controlled and those not controlled by Ir genes: both types involve the recognition of the antigen, in the context of Mhc molecules.Abbreviations used in this paper A class II MHC molecule consisting of the A and A chains - ADH alcohol dehydrogenase - APC antigen-presenting cell - cpm counts per minute - E class II Mhc molecule onsisting of the E and A chains - Ir immune response - KLH key-hole limpet hemocyanin - Mhc major histocompatibility complex - PBS phosphate buffered salt solution - SD standard deviation - SI stimulation index - SN supernatant of Sendai-virus preparation     

Plasmid-Mediated Gene Transfer Between Insect-Resident Bacteria, Enterobacter cloacae, and Plant-Epiphytic Bacteria, Erwinia herbicola, in Guts of Silkworm Larvae

Five strains of Enterobacter cloacae isolated from several species of plants and insects were able to grow in the guts of silkworm larvae. A much larger population of Ent. cloacae strains was detected in the insect guts and feces collected 3 and 6 days than in samples collected 1 day after feeding artificial diets contaminating these bacteria. Furthermore, insect-origin strains of Ent. cloacae were mated with a donor strain, epiphytic Erwinia herbicola, harboring RSF1010 and pBPW1::Tn7 plasmids in the insect guts by introducing these bacteria through separate artificial diets administered at different times. A number of transconjugants, Ent. cloacae strains which had acquired RSF1010 plasmid, were detected from guts and fecal samples at transfer frequencies of 10⁻² to 10⁻³ per recipient. Thus, gene transfer between epiphytic Er. herbicola and insect-resident Ent. cloacae strains in the insect guts was confirmed. These findings may provide significant information about the role of ′′in insecta mating' in the evolution of these bacteria. Received: 16 June 1998 / Accepted: 7 July 1998     

Antigenic characterization of Microsporum canis,M. distortum,M. equinum,M. ferrugineum and Trichophyton soudanense cultures

The authors investigated the antigenic properties of a group of closely related dermatophytes whose components may be morphologically confused with one another: Microsporum canis, M. distortum, M. equinum, M. ferrugineum and Trichophyton soudanense. By using reference antigens and adsorbed monospecific antisera, it was possible to serologically distinguish the reference strains by the exoantigen technique. Their common and specific antigenic determinants were visualized by cross-immunoelectrophoresis tests with intermediate gel.     

Morphological characteristics and phylogenetic relationship of <Emphasis Type="Italic">Anabaena</Emphasis> species from Lakes Dianchi and Erhai,China

Although Anabaena is one of the most prevalent planktonic freshwater genus in China, there are few taxonomic reports of Anabaena strains by morphology and genetics. In this study, morphological characteristics and phylogenetic relationships of seven Anabaena strains isolated from two plateau lakes, Lakes Dianchi and Erhai, were investigated. Morphological characteristics such as morphology of filament, cellular shapes and sizes, relative position of heterocytes and akinetes, and presence or absence of aerotopes, were described for these seven strains. Phylogenetic relationships were determined by constructing 16S rRNA gene tree using the neighbor-joining algorithm. The seven strains were morphologically identified as three groups, and phylogenetic analysis based on 16S rRNA gene sequences also showed that these seven strains were in three groups. Strains EH-2, EH-3, and EH-4 were in group A belonging to the Anabaena circinalis and A. crassa group, and strains DC-1, DC-2, and EH-1 were in group B and identified as A. flos-aquae. Strain DC-3 without aerotopes was significantly different from the other isolated strains and was determined as A. cylindrica. Handling editor: J. Padisak     

Specificity of cell-mediated transplantation reactions in rats detected by the macrophage adherence inhibition test

The macrophage adherence inhibition (MAI) test, previously described as a correlative to specific cellular immunity, has been used to study the specificity of primary cell reactions after allografting in rats. The adherence of sensitized peritoneal cells (PCs) from AVN rats (major histocompatibility haplotypeH-1 ^a ) bearing Lewis skin grafts (H-1 ^l ) was inhibited specifically by antigens from the graft donor and by those of the allogeneic strains BN (H-1 ⁿ ), BD V (H-1 ^d ), and DA (H-1 ^a ). Antigens of strain AVN, congenic strain Lewis. 1A (H-1 ^a ), and xenoantigens did not inhibit adherence. In general, positive reactions occurred during interaction of the sensitized PC with antigenic material sharing some components of theH-1 system with the graft donor. On the other hand, no crossreactions between individual rat strains were found when cytotoxic antibodies were tested after skin allografting. The negative MAI test in PC from AVN rats sensitized with skin allografts from the Lewis strain done with Lewis. 1A (H-1 ^a ) antigenic material, and the positive MAI test with antigenic material of the DA (H-1 ^a ) strain point to the possible existence of serologically undefined transplantation antigens participating in the cell-mediated reactions.     

The Content of Antigens 1, 2 and 3 in Strains of Bordetella pertussis and in Vaccines

A comparison was made of the content of heat-labile antigens 1, 2 and 3 in strains of Bordetella pertussis isolated in Toronto, and the content of the same antigens in vaccines being used in that community. Antisera were prepared in rabbits and made monospecific by absorptions. Antigens were detected by slide agglutination tests. Of the 58 strains examined, 56 contained antigens 1 and 3 but no antigen 2 was detected. Nine of these strains were isolated from children who had received a full course of vaccination and had subsequently developed pertussis. All 10 vaccines examined contained antigen 1; two, possibly four, contained no antigen 2; and seven of the 10 vaccines were weak or deficient in antigen 3. This work suggests that improved protection might be attained by the use of vaccines containing adequate amounts of all three antigens.     

Genetic control of acquired resistance to visceral Leishmaniasis in mice

A series ofH-2 and non-H-2 congenic resistant (CR) strains on a C57BL/10Sn background were infected with 10⁷ amastigotes ofLeishmania donovani. Non-H-2 congenic strains B10.LP-H-3 ^b and B10.CE(30NX) and (B10.LP-H-3 ^b × B10)F₁ hybrids showed a very rapid decrease in liver-parasite burdens beyond day 21. Parasite counts for these strains at day 35 were significantly lower than for all other strains tested. The rapid decrease in parasite numbers, massive lymphocellular infiltration into the liver and strong delayed hypersensitivity reactions to parasite antigens in strains congenic for a portion of chromosome 2 indicated that acquired immunity toL. donovani was controlled by a dominant gene at or near theIr-2 locus. In addition, B10.129(10M) mice, which differ from C57BL/10Sn at theH-11 locus, showed highly significant increases in parasite numbers at day 35. Other observations supporting the absence of acquired immunity in B10.129(10M) included negative delayed hypersensitivity tests to parasite antigens and the absence of lymphocellular infiltrate into the liver. Although the differences were not as pronounced,H-2 CR strains withH-2 ^b ,H-2 ^a , andH-2 ^k haplotypes also showed significantly greater decreases in parasite numbers by day 35 as compared to otherH-2 CR strains.     

Identification of new murine lymphocyte alloantigens: Antisera prepared between C57L, 129 and related strains define new loci

Antisera prepared by immunizing between the strains 129 and C57L and other related strains identified new antigens expressed on lymphocytes and in particular on thymocytes. Absorption analysis demonstrated that the antisera were complex, and contained several new antibodies including some which were not cytotoxic, but could be detected by rosetting. The loci defined by these antibodies are referred to asLy-9, Ly-11, Ly-12, Ly-13, andLy-14, although several of the antigens were not confined to lymphocytes. In addition, the Ea-7 specificities, previously considered to be purely red-cell alloantigens, were also found on thymocytes.