Prevalence of yersinia plasmid-encoded outer protein (Yop) class-specific antibodies in multitransfused Greek patients with thalassemic syndromes

Objective: To evaluate the prevalence of class-specific antibodies (G, A, M) to Yersinia enterocolitica plasmid-encoded outer proteins (Yops), in a closely followed multitransfused population of patients with thalassemia.
Methods: Sera from 408 β-thalassemic patients and 386 healthy blood donors used as controls were analyzed with the enzyme-linked immunosorbent assay (ELISA) for IgG, IgA and IgM antibodies to yersinia outer proteins. The Yop antigen for the ELISA was prepared using a plasmid-bearing wild-type strain of Y. enterocolitica of serotype 0:8.
Results: Anti-Yop IgG antibodies were detected in 84 out of 408 β-thalassemic patients (20.6%) compared with only eight out of 386 (2.1%) healthy blood donors. None of the sera of either group was positive for anti-Yop IgA or IgM antibodies. On evaluating patients with registered clinical and laboratory signs of a previous yersinia infection in the period from 1978 to 1996, we found that those with a positive aggiutination test for Y. enterocolitica infection at the time of manifestation showed a higher rate of persisting IgG seropositivity to Yops than those with positive culture and clinical signs only. A significant percentage (9.49%) of the seropositive patients had no registered data of a past Y. enterocolitica infection. There was remarkable persistence of anti-Yop IgG antibodies in the thalassemic population, even in patients infected during the early years of our study period (1978–80).
Conclusions: The results suggest that the determination of class-specific antibodies to Yops, which are specific antigens for the pathogenic yersiniae ( Y. enterocolitica, Y. pseudotuberculosis and Y. pestis ), in addition to its usefulness in the diagnosis of infection, will be a very sensitive and specific index for epidemiologic studies.     

Yersinia enterocolitica O:9 as a potential live oral carrier for protective antigens

Yersinia enterocolitica has the capacity to invade the intestinal tissue and to resist the primary host resistance. The former is chromosome coded while the second largely depends on the presence of a 70 kb plasmid called pYV. This plasmid directs the conditional synthesis of high amounts of proteins (YOPs) that are secreted and inserted in the outer membrane. In order to evaluate Y. enterocolitica W22703 as a potential live carrier for immunization, three strains expressing beta-galactosidase (GZ), were tested for their ability to induce an antibody response to this antigen in mice. The first strain contained plasmid pGC1256, a mutated pYV plasmid containing lacZ transcribed from a yop gene promoter. This strain produced high amounts of GZ instead of a YOP protein and was shown to be hypovirulent. The other strains tested were W22703 pYV+ and pYV- containing a derepressed lac operon carried on an independent plasmid. Immunoblot analysis of sera of mice having received by oral inoculation, W22703(pGC1256) or the pYV+ GZ producing strain revealed the presence of antibodies to GZ. The response to GZ after inoculation of W22703(pGC1256) was shown by ELISA to be only slightly inferior to that obtained by subcutaneous injection of GZ. No response was obtained after oral inoculation of the pYV-GZ producing strain. This showed that the presence of pYV was necessary to obtain an antibody response in this system.     

Analysis of the class-specific immune response to Yersinia enterocolitica virulence-associated antigens in oro-gastrically infected rabbits

The antibody response was analysed in rabbits oro-gastrically infected (i) with virulent-(plasmid-carrying) Yersinia enterocolitica 0:3 (v-rabbits) and (ii) with the avirulent (plasmid-cured) derivative (av-rabbits). In an immunoblot assay with whole cell lysate proteins from the infecting virulent Yersinia strain, a significant IgG response was evident in convalescent-sera of v-rabbits and of av-rabbits, demonstrating that all rabbits seroconverted. However, v-rabbits mounted a stronger immune response to the cell lysate proteins than av-rabbits and the immune response persisted for a longer time. The post-challenge sera also reacted with whole cell lysate proteins of Escherichia coli and Salmonella typhimurium, indicating cross-reactivity between the different members of Enterobacteriaceae. In contrast, the antibody response against plasmid-encoded released proteins (RPs) appeared specific for infection with virulent strains in that the sera of av-rabbits failed to recognize the plasmid-encoded proteins. Six days following challenge with the virulent Yersinia strain all animals mounted a serum IgM and IgA response to RPs, followed by IgG antibodies on day 9. While the IgM and IgA serum antibody response rapidly decreased (within five-seven and eight-ten months, respectively), IgG antibodies to RPs were still present one year after challenge. Fourteen months after the first infection both the av-rabbits and the v-rabbits were reinfected with the virulent Yersinia strain and the antibody response to RPs was monitored. The v-rabbits only responded with a significant increase of IgG antibodies, indicating that they were primed to the RPs whereas the av-rabbits produced IgM, IgA and IgG specific antibodies like those seen in the primary response of v-rabbits. This study indicates that the rabbit model is helpful and adequate to analyse the character and kinetics of the antibody response during Yersinia infection.     

Adhesion of Yersinia enterocolitica to human epithelial cell lines and to rabbit and human small intestinal tissue

In assays to determine adherence of Yersinia enterocolitica, virulence plasmid-containing (pYV+) strains and strains of their plasmid-cured (pYV-) derivatives adhered equally efficiently to HeLa cells and fetal intestinal epithelial INT 407 cells. Non-pathogenic strains of Y. enterocolitica did not adhere. In contrast, pYV+ strains adhered more efficiently to rabbit and human intestine than did pYV(-)-strains, as evaluated by an in vitro assay, measuring adhesion of radiolabeled bacteria to disks of intestinal tissue. Thus, even if pathogenic strains of Y. enterocolitica are able to adhere to cultured epithelial cell lines and intestinal tissue by means of chromosome-encoded properties alone, properties encoded by the pYV plasmid significantly enhance and contribute to adhesion to intestinal tissue.     

Serum immunoglobulin levels in Australia antigen positive and Australia antigen negative hepatitis

Ig levels were determined by radial immunodiffusion in uncomplicated cases of acute hepatitis with or without Australia antigenaemia. Initial sera from Australia antigen negative cases showed a striking elevation in IgM levels when compared to Australia antigen positive cases (6·5 versus 1·9 mg/ml). None of twenty-four Australia antigen positive cases exceeded 3 mg/ml IgM, and only 3/58 Australia antigen negative cases exhibited values below 3 mg/ml. Intial sera from Australia antigen positive and Australia antigen negative subjects did not differ in concentration of IgG, IgA, or IgD. Serial determinations of IgG revealed a transient fall in patients with Australia antigen positive hepatitis, and a rise in Australia antigen negative cases. Asymptomatic, Australia antigen positive, Guaymi Indian subjects were compared to matched Australia antigen negative controls from the same indigenous group and no differences in the concentration of IgG, IgM, IgA or IgD were found, although elevations of IgG and IgM were common in both groups. No evidence of abnormal proteins was found when sera were tested by cellulose acetate electrophoresis or by immunoelectrophoresis versus immunoglobulin-specific antisera. Ultracentrifugal analysis failed to detect `7S' IgM.     

Use of protein A-treated sera in unmasking herpes simplex virus type 1 (HSV-1) immunoglobulin A and identifying HSV-1 immunoglobulin G as the predominant neutralizing antibody.

Treatment of human sera with protein A reduced the amounts of immunoglobulin G (IgG), IgA, and IgM detected by radial immunodiffusion. This treatment also decreased the amount of herpes-specific IgG and IgM detected by radioimmunoassay, whereas it increased and even unmasked the amount of herpes-specific IgA detected. Comparison of protein A-treated sera with untreated sera indicated that herpes simplex virus type 1 IgG was responsible for more than 92 to 99% of the serum neutralizing activity.     

Immune response in dogs experimentally infected with Paracoccidioides brasiliensis.

The aim of the present study was to evaluate the immune response of young dogs experimentally infected with Paracoccidioides brasiliensis. Six dogs were infected intravenously with P. brasiliensis and one control dog was inoculated with sterile saline. The infected animals were sacrificed in groups of two at 1, 6 and 12 months after infection. During the experimental period, the immune responses of the dogs to the fungus were followed by ELISA (IgM and IgG), by the immunodiffusion test and by the skin test with gp43. After killing the dogs, samples from several organs were submitted to histopathological analysis (H&E and Grocott stains) but the fungus was not observed in any tissue. Attempts to isolate the fungus from these tissue samples were also unsuccessful. All infected dogs, except one, reacted positively to the immunodiffusion and skin tests. All infected dogs showed a humoral immune response to the gp43 antigen detected by ELISA. The IgM and IgG response peaked by the first and second month, respectively. We conclude that young dogs appear to be resistant to the development of paracoccidioidomycosis.     

The influence of properties encoded by the Yersinia virulence plasmid on adhesion of Yersinia enterocolitica to ileal brush border membrane vesicles

The influence of plasmid-associated cell surface structures on the ability of Yersinia enterocolitica to bind to ileal brush border membrane vesicles (BBVs) was investigated. Rabbit or human BBVs were immobilized on polystyrene microtiter plates and adhesion of radiolabeled cells of Y. enterocolitica was determined. Strains of pathogenic serotypes carrying the Yersinia virulence plasmid (pYV+), as well as their isogenic plasmid cured derivatives (pYV-), adhered to immobilized BBVs, but adhesion of pYV+ organisms was markedly greater than that of pYV- ones. Strains belonging to non-pathogenic serotypes did not adhere significantly. The pYV+ strain Ye0301P+ did not express specific adhesion to glycolipids, nor was adhesion to BBVs reduced in the presence of various monosaccharides. Proteolytic digestion of surface structures on strain Ye0301P+ markedly reduced adhesion. pYV+ strains also demonstrated greater adhesion to a non-biological surface (polystyrene) and showed a higher degree of hydrophobicity than pYV- organisms as evaluated in a two-phase partitioning system. It is therefore likely that the plasmid-associated adhesion of Y. enterocolitica is promoted by one or more outer membrane proteins, that confer hydrophobicity to the bacterial surface.     

Comparison of crossed immunoelectrophoresis, enzyme-linked immunosorbent assays, and tube agglutination for serodiagnosis of Yersinia enterocolitica serotype O:3 infection.

Antibodies against Yersinia enterocolitica serotype O:3 were measured by crossed immunoelectrophoresis (XIE) using whole-cell sonic extract as antigen and by enzyme-linked immunosorbent assays (ELISAs) using either purified lipopolysaccharide or whole formalinized cells expressing virulence plasmid-encoded surface antigens (pYV+ cells). The results were compared with those obtained with the standard tube agglutination method. Sera from three groups of people were examined by using these assays. The first group consisted of healthy blood donors, the second consisted of patients with recent infection due to microorganisms other than Y. enterocolitica O:3, and the third consisted of patients with recent Y. enterocolitica O:3 infection. Sera from the last group were also obtained at regular intervals for 12 months postinfection. Results obtained with XIE and the ELISAs were in good agreement with those obtained with tube agglutination. Variation, diagnostic sensitivity, and diagnostic specificity were satisfactory for all the assays studied. However, the lipopolysaccharide ELISA was less laborious than tube agglutination and XIE and carried a somewhat greater diagnostic specificity than the pYV+ ELISA. XIE and the pYV+ ELISA, on the other hand, also had advantages. XIE enabled simultaneous examination of the individual antibody response against a wide range of chromosome-encoded antigens, and the pYV+ ELISA enabled detection of specific pYV antibodies when sera were adsorbed with formalinized pYV-cured Y. enterocolitica O:3 cells prior to the assay.     

Antibody responses to Campylobacter infections determined by an enzyme-linked immunosorbent assay: 2-year follow-up study of 210 patients

An enzyme-linked immunosorbent assay (ELISA) was adapted to measure immunoglobulin G (IgG), IgM, and IgA classes of human serum antibody to Campylobacter jejuni and Campylobacter coli. Heat-stable antigen, a combination of C. jejuni serotype O:1,44 and O:53 in the ratio 1:1, was used as a coating antigen in the ELISA test. A total of 631 sera from 210 patients with verified Campylobacter enteritis were examined at various intervals after infection, and a control group of 164 sera were tested to determine the cut-off for negative results. With a 90th percentile of specificity, IgG, IgM, and IgA showed a sensitivity of 71, 60, and 80%, respectively. By combining all three antibody classes, the sensitivity was 92% within 35 days after infection, whereas within 90 days after infection, a combined sensitivity of 90% was found (IgG 68%, IgM 52%, and IgA 76%). At follow-up of the patients, IgG antibodies were elevated 4.5 months after infection but exhibited a large degree of variation in antibody decay profiles. IgA and IgM antibodies were elevated during the acute phase of infection (up to 2 months from onset of infection). The antibody response did not depend on Campylobacter species or C. jejuni serotype, with the important exception of response to C. jejuni O:19, the serotype most frequently associated with Guillain-Barré syndrome. All of the patients infected with this serotype had higher levels of both IgM (P = 0.006) and IgA (P = 0.06) compared with other C. jejuni and C. coli serotypes.     

Evaluation of serological tests for diagnosis of Brucella melitensis infection of goats.

Five serological assays were evaluated for the diagnosis of brucellosis in goats: the rose bengal test (RBT), complement fixation test (CFT), radial immunodiffusion (RID) with Brucella and Yersinia enterocolitica O:9 polysaccharides, counterimmunoelectrophoresis (CIEP) with cytosol, and enzyme-linked immunosorbent assay (ELISA) with polyclonal and protein G conjugates and smooth lipopolysaccharide (S-LPS), native hapten polysaccharide (NH), or cytosol antigens. For optimal sensitivity, RBT had to be used with sera-antigen at a 3:1 dilution. In the RID test, Brucella melitensis biotype 1 NH could not be replaced by Brucella abortus biotype 1 or Y. enterocolitica 0:9 polysaccharides. In the ELISA, S-LPS and NH gave similar results and the protein G conjugate increased the specificity. With the sera from 55 B. melitensis culture-positive goats, the sensitivity was 100% for RBT, CFT (titer > or = 4), and ELISA with S-LPS or NH; 94% for RID; and 93% for CIEP. All tests were negative (100% specific) when testing the sera from 127 brucella-free goats. Larger discrepancies among the results of the serological tests were obtained with sera from goats of areas where brucellosis is endemic. When the sera of 20 young goats vaccinated subcutaneously (10(9) CFU of B. melitensis Rev 1) and bled 6 months later were examined, the specificities were as follows: NH ELISA, 60%; CFT and S-LPS ELISA, 75%; RBT, 80%; CIEP, 90%; and RID, 94%. With the sera from 10 young goats vaccinated conjunctivally (10(9) CFU of B. melitensis Rev 1) all tests were 100% specific 4 months after vaccination. The proportion of goats giving a positive reaction after vaccination decreased faster in RID than in other tests.     

日本血吸虫成虫及虫卵可溶性抗原的早期诊断分子筛选

为获取有效的日本血吸虫病早期诊断靶抗原分子。以感染前和感染后2周、4周、6周兔混合血清以及急性、慢性日本血吸虫病人和正常人血清,用Westernblot对日本血吸虫成虫可溶性抗原(AWA)和虫卵可溶性抗原(SEA)进行了全面的分析与筛选。SEA140kDa和AWA54kDa、21kDa、20kDa分子出现最早,能被感染后2周兔血清所识别;SEA69kDa、50kDa、45kDa、38kDa和AWA72kDa、45kDa、34kDa、15kDa相继能被感染后4周兔血清所识别;其中SEA140kDa、50kDa、45kDa、38kDa和AWA34kDa、21kDa、54kDa、15kDa与病人血清反应同6周兔血清反应效果相仿,均为免疫反应主带,且在SDS-PAGE上有对应的蛋白主带,具有潜在的早期诊断价值。进一步对SEA进行抗体类型的分析,发现SEA能刺激机体产生IgG、IgM和IgA反应,其中SEA50kDa、45kDa、38kDa三个抗原分子均能被IgG、IgM、IgA三种抗体所识别,且均为反应主带;SEA140kDa以IgM反应带最深;SEA100kDa反应带仅出现于病人血清,以IgM反应最强,且其与IgM反应带在急性病人血清明显深于慢性病人血清,表明针对IgM的SEA100kDa分子也具有一定的早期诊断和区分病程的价值。SEA140kDa、50kDa、45kDa、38kDa和AWA34kDa、21kDa、54kDa、15kDa和针对IgM的SEA100kDa分子是具有潜在早期诊断价值的优势靶抗原分子。     

Class-specific antibody determination against Aspergillus fumigatus by means of the enzyme-linked immunosorbent assay. III. Comparative study: IgG, IgA, IgM ELISA titers, precipitating antibodies and IgE binding after fractionation of the antigen

IgG, IgA and IgM ELISA antibody titers against Aspergillus fumigatus were elevated in sera of patients with aspergilloma and allergic bronchopulmonary aspergillosis (ABPA), showing higher titers for the IgG antibodies compared with the IgA and IgM antibodies. No differences were found between titers of identical antibody classes in the two groups of sera. IgG and IgA ELISA titers were highly specific whereas IgM ELISA showed more unspecific binding of IgM antibodies. Antibodies, as measured by ELISA, studied after fractionation of the antigen into fractions of decreasing molecular weight, showed a preferential binding by the high molecular weight fractions. Precipitating antibodies studied in patient sera did not always correspond with the IgG ELISA titers. IgE antibody binding was observed in all fractions from Sephadex G-100 fractionated components; maximum binding was found with fractions of 28,000-60,000 daltons. The low molecular weight fractions (18,000-less than 5,000 daltons) showed less IgE binding but the quantity of this fraction was higher. The discrepancies noted between the IgG and IgA ELISA titers and the binding of IgM or IgE antibodies indicate that antigenic components may in part differ in the binding of antibody classes.     

Immune response to plasmid- and chromosome-encoded Yersinia antigens.

The immune response of humans and mice to temperature-specific, plasmid- or chromosome-encoded proteins of yersinia pestis and Yersinia enterocolitica was investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Extracts from Y. pestis and Y. enterocolitica strains with and without the virulence plasmids pYV019 and pYV8081, respectively, were resolved by denaturing electrophoresis, and the major antigens were visualized with sera from convalescing plague patients, individuals immunized with plague vaccine, and mice and rabbits immunized with avirulent live yersiniae. The Y. pestis grown in vitro in this study did not express detectable amounts of plasmid-encoded antigens. The sera from plague patients recognized Y. pestis and Y. enterocolitica antigens ranging from 15 to 72 kilodaltons (kDa), whereas sera from immunized subjects recognized four antigenic components in Y. pestis ranging from 17 to 64 kDa and five antigens in Y. enterocolitica ranging from 16 to 68 kDa. Sera from mice reacted with 7 antigens in Y. pestis and 12 antigens in Y. enterocolitica ranging from 14 to 68 kDa, and sera from rabbits reacted with 7 and 10 antigens in Y. pestis and Y. enterocolitica, respectively. All of the plague patient sera, as well as the sera from immunized mice and rabbits, reacted with a 22-kDa Y. enterocolitica plasmid-associated polypeptide, and five of the patient sera also recognized a Y. enterocolitica plasmid-associated 31-kDa protein. The results indicate a common immune response to at least these two plasmid-specified Yersinia outer membrane proteins. Y. pestis apparently expresses these components only in vivo, and in vitro, Y. enterocolitica expresses a greater number of plasmid-associated antigens than does Y. pestis.     

HC-IgA antibodies of different specificities are normally present in serum: quantitation by ELISA and relationship to the major Ig classes.

Enzyme-linked immunosorbent assays (ELISA) were developed for direct measurement of protein HC-IgA complexes (HC-IgA) in serum with antibody specificity for rabbit IgG (rheumatoid factor (RF) activity), lipopolysaccharide from Yersinia enterocolitica serotype O:3 (Y3) and tetanus toxoid (TT). About 80% of patients with rheumatoid arthritis had increased concentrations of HC-IgA-RF. The values were correlated with the concentrations of IgA-RF and IgM-RF. HC-IgA anti-Y3 was measured in 45 sera with anti-Y3 antibodies of IgM, IgG and IgA class. The HC-IgA anti-Y3 levels were correlated with those of anti-Y3 of IgG and IgM class, but not of IgA class. For HC-IgA anti-Y3, the closest correlation was that with the specific IgM antibody concentration, rs = 0.63 (p less than 0.001). In 25 normal sera, significant correlations were observed between HC-IgA anti-TT and specific antibodies of IgG and IgA class, but not of IgM class. In 107 sera containing IgA M-components, the total concentration of HC-IgA correlated poorly with both protein HC and with IgA concentrations. It was concluded that specific HC-IgA antibodies are normal constituents of serum, and that their concentrations are not directly related to the serum content of specific IgA antibodies.     

Interaction of Yersinia enterocolitica and Y. pseudotuberculosis with platelets.

Yersinia enterocolitica is a bacterium capable of growth at 4 degrees C in donated blood and has been responsible for many deaths following transfusion. Interaction of Y. enterocolitica with blood cells is of interest in understanding the mechanisms of survival and growth in blood. The closely related organism Y. pseudotuberculosis is known to invade platelets and cause platelet aggregation by a mechanism that involves expression of the chromosomal inv gene. Yersinia isolates were made to express green fluorescent protein (GFP) and their interaction with platelets was studied by flow cytometry, enterocolitica did not cause platelet aggregation or activation, not even when grown at 22 degrees C to maximise inv expression. Attachment of Y. enterocolitica O:9 to platelets occurred with virulence plasmid-bearing (pYV+) strains grown at 37 degrees C but not with pYV- strains nor with strains grown at 22 degrees C. Y. pseudotuberculosis containing inv did cause platelet activation and aggregation when grown at 22 degrees C, as has been shown before, but also showed enhanced attachment to platelets when grown at 37 degrees C. Electron microscopy studies confirmed that inv-expressing Y. pseudotuberculosis invaded platelets but Y. enterocolitica attached only to the outer surface of platelets. Interaction of Y. enterocolitica O:9 with platelets provided a modest protection against bacterial killing by human serum. Interaction of Y. enterocolitica O:9 with platelets does not lead to platelet invasion or activation, and is mediated through plasmid-coded factors, not inv.     

Comparative tests for detection of plague antigen and antibody in experimentally infected wild rodents.

The enzyme-linked immunosorbent assay (ELISA) was compared with other standard tests for detection of plague (Yersinia pestis) antibody and antigen in multimammate mice (Mastomys coucha and M. natalensis) which were experimentally infected and then killed at daily intervals postinoculation. For detection of antibody in sera from M. natalensis, the immunoglobulin G (IgG) ELISA was equivalent in sensitivity to passive hemagglutination and more sensitive than the IgM ELISA and complement fixation. Antibody was first detected on postinfection day 6 by all four tests, but IgM ELISA titers had declined to undetectable levels after 8 weeks. For detection of fraction 1 Y. pestis antigen in rodent organs, the ELISA was less sensitive than fluorescent antibody but more sensitive than complement fixation or immunodiffusion. Plague fraction 1 antigen was detected in 16 of 34 bacteremic sera from M. coucha and M. natalensis. The threshold sensitivity of the ELISA was approximately 10(5) Y. pestis per ml.     

Determination of IgA antibodies to human cytomegalovirus by enzyme-linked immunosorbent assay (ELISA)

A sensitive enzyme-linked immunosorbent assay (ELISA) is described for detection of IgA antibodies cytomegalovirus (CMV). The antigen consisted of a sonicated extract of CMV infected human embryo cells. The tested sera were absorbed with staphylococcus aureus (strain Cowan 1) before analysis. Rabbit antihuman IgA peroxidase conjugate was used to detect human IgA bound to viral antigen. In parallel, Igm and IgG antibodies to CMV were studied by ELISA and by the immunoperoxidase antibody to membrane antigen (IPAMA) technique, respectively. CMV IgA was detected in high titers by ELISA in eight of nine CMV patients. The maximal IgA titers were generally lower than the IgM titers detected by ELISA. No CMV IgA antibodies (titer less than 20) were detected by ELISA in 57 control sera (healthy adults, hospitalized patients with various other diseases), paired sera of five patients with acute herpes simplex, infection, two patients with Epstein-Barr infections, one patients with varicella, and one with zoster infections. The potential application of the ELISA CMV IgA technique in serodiagnosis of CMV infections is discussed.     

Humoral immune response in Epstein-Barr virus infections. I. Elevated serum concentration of the IgG1 subclass in infectious mononucleosis and nasopharyngeal carcinoma

Using radial immunodiffusion we measured IgG subclass concentrations and studied their distribution in serum samples from patients with infectious mononucleosis (IM) and nasopharyngeal carcinoma (NPC), two Epstein-Barr virus (EBV)-associated diseases, in comparison with two control groups [completely anti-EBV negative persons and subjects carrying antibodies to the viral capsid antigen (VCA)]. Antibody titres to VCA and to the early antigen (EA) were determined by indirect immunofluorescence and revealed characteristic patterns for the respective diagnostic groups. Nephelometric assays served for quantitating total protein, albumin, total IgG, IgA and IgM in all the sera. In the IM and NPC groups the concentration of IgG1 was significantly elevated by more than 50% whereas the other three subclasses remained unchanged as compared with the controls. Correspondingly, we found a significant increase of total IgG in IM and NPC. In IM, the only disease where VCA-specific IgM antibodies have been reported to occur, IgM levels were markedly elevated. Our data suggest that the IgG1 subclass plays an important role in the humoral immune response to EBV-determined antigens and that it is possibly involved in the control of virus infection.     

Immunoglobulins A, G, and M in serum and in some secretions of monkeys (Macaca fascicularis syn. irus).

The purpose of this investigation was to study the distribution and levels of the following immunoglobulins, IgA, IgG, and IgM ,in sera and in some secretions of monkeys (M. fascicularis). IgG, IgA, and IgM were isolated from monkey serum and secretory IgA was separated from monkey milk by combined gel filtration and ion-exchange chromatography. These pure preparations served as standards to quantitate immunoglobulins in sera and secretions by single radial immunodiffusion. Antisera were raised in the rabbit against the pure immunoglobulins and also against the whole secretions to identify the immunoglobulins in immunoelectrophoresis. In common with humans, the major immunoglobulin in serum and amniotic fluid is IgG and the IgG/IgA ratio is greater than unity. In secretions IgA is the dominant immunoglobulin and the IgG/IgA ratio is less than 1. In general, the levels of immunoglobulins in the sera and secretions of monkeys paralleled the levels found in humans. No age-related increase in immunoglobulin levels was detected in the sera of monkeys.