Comparative metabolism and disposition of gemfibrozil in male and female Sprague-Dawley rats and Syrian golden hamsters

Antibodies (Abs) can contribute to the cure of a viral infection, in principle, in two ways by: (1) binding to infected cells and thereby reducing the production of progeny virus [here termed cell-targeting (CT) activity] and (2) reacting with released progeny virus and thereby inhibiting the spread of the infection [termed virus neutralizing (VN) activity]. We have previously shown that a pulmonary influenza virus infection in severe combined immunodeficient mice could be cured by treatment of these mice with hemagglutinin (HA)-specific monoclonal Abs (mAbs) that mediated both of the above activities. Although the therapeutic activity of these mAbs correlated with their VN activity, it remained unclear how much their CT activity contributed to the Ab-mediated recovery process. To clarify this point, we tested the therapeutic efficacy of two mAbs of IgG2a isotype that mediated CT but no VN activity: one specific for the viral neuraminidase and the other for matrix protein 2. Both mAbs reduced pulmonary virus titers by 100- to 1000-fold but they failed to clear the infection, even when administered in combination and at therapeutically saturating concentrations. The results suggest that CT activity contributes significantly also to the therapeutic activity of HA-specific mAbs and further support the notion that VN-activity is required for Ab-mediated virus clearance.     

Human immune responses to influenza virus vaccines administered by systemic or mucosal routes

Healthy adult volunteers were immunized by parenteral or oral routes with trivalent inactivated influenza vaccine (A/Chile/1/83 (H1N1), A/Mississippi/1/85 (H3N2), and B/Ann Arbor/1/86), or intranasally with live attenuated, cold-adapted influenza type A/Texas/1/85 (H1N1) reassortant virus. In all volunteers, cells spontaneously secreting IgA, IgG or IgM antibodies specific to influenza virus were detected in peripheral blood on days 6-13 after immunization, and specific IgA, IgG and IgM antibodies to influenza vaccine were measured in sera and external secretions (saliva and nasal lavage). Following systemic immunization, a raise in specific antibodies of all isotypes was observed in sera beginning on day 13. Although small variations in IgA and IgM antibodies in saliva and nasal lavages were detected, antigen-specific IgG significantly increased between days 13 and 27. Intranasal administration of attenuated virus induced IgA and IgG antibodies in serum as well as in secretions. Serum antibodies were not substantially influenced by oral immunization, only a small increase in all isotypes was observed in volunteers' sera 21 days after ingestion of vaccine. However, in secretions, antigen-specific IgA and IgG responses were detected one week after immunization and reached a peak response on day 20. These studies show that different routes of immunization can be effective for the induction of specific antibodies, and support the concept of the common mucosal immune system in humans by demonstrating that the oral or intranasal administration of antigen-induced specific antibodies of IgA isotype in external secretions, preceded by the transient appearance in peripheral blood of specific antibody-producing cells.     

Isotype profiles of Leishmania donovani-infected BALB/c mice: preferential stimulation of IgG2a/b by liposome-associated promastigote antigens

Leishmaniasis presents a complex spectrum of diseases and immunological manifestations depending upon both the species of the microorganism and the host it infects. BALB/c mice, which are homozygous for Lsh(s) on chromosome 1, are genetically susceptible to the visceralizing species of Leishmania. Infection of these mice with an Indian strain of Leishmania donovani showed a steady rise in the level of parasite burden in both the liver and the spleen to 24 wk. To investigate the immune responses determining the course of infection, we studied the relative levels of specific IgG, IgM, and IgA antibodies, and IgG isotypes, in the sera of diseased and protectively immunized mice at different periods of infection. IgG1 and IgG2a were stimulated in the control, infected, and immunized mice after parasite challenge. However, an early induction of IgG1 in the normal infected mice and stimulation of IgG2a and IgG2b isotypes prior to parasite challenge in liposome-antigen-immunized mice suggest that the elicitation of a particular subset of CD4+ T cells at the onset of disease may be responsible for either progression or resolution of infection.     

Human and nonhuman primate lymphocytes engrafted into SCID mice reside in unique mesenteric lymphoid structures

The present study compares the location and phenotype of B lineage lymphocytes in tissues from SCID mice engrafted with PBMC of human, chimpanzee, and pig-tailed macaque origin. In mice repopulated with both human and nonhuman primate lymphocytes, plasma cells were found in the peritoneal cavity in vascularized structures located in the mesentery near the pancreas, intestines, and spleen. The predominant isotype of the plasma cells was IgG; IgM and IgA cells were also present. Kappa and lambda light chains were expressed by 62% and 38% of the Ig-containing cells, respectively. J chain expression occurred in most cells irrespective of the Ig isotype. In the SCID mice engrafted with human lymphocytes, a few IgM-containing cells were found in the spleen; plasma cells were not found in other tissues, including the intestine. The aggregation of plasma cells did not appear to be a result of infection with EBV. T cells were rarely found in the lymphoid aggregates but were recovered from the spleen and peritoneal lavage. Human Ig levels in the serum of engrafted mice reflected the isotype distribution of the cells with IgG > IgM > or = IgA.     

In vivo anti-influenza virus activity of Kampo (Japanese herbal) medicine "sho-seiryu-to"--stimulation of mucosal immune system and effect on allergic pulmonary inflammation model mice

When BALB/c mice were treated with a Kampo (Japanese herbal) medicine "Sho-seiryu-to (SST)" (1 g/kg, 10 times) orally from 7 days before to 5 days after the infection and infected with mouse-adapted influenza virus A/PR/8/34 by nasal-site restricted infection, SST caused increment of the influenza virus hemagglutinin-specific IgA antibody secreting cells in nasal lymphocyte but not in Peyer's patch lymphocyte at 6 days after infection in comparison with water-treated mice. Oral administration of SST also augmented IL-2 receptor beta chain+ (activated) T-cell in Peyer's patch lymphocyte, but not in the nasal lymphocyte. We previously reported that SST showed potent anti-influenza virus activity through augmentation of the antiviral IgA antibody titer in the nasal and broncho-alveolar cavities of the mice (T. Nagai and H. Yamada, 1994, Int. J. Immunopharmacol. 16, 605-613). These results suggest that oral administration of SST shows anti-influenza virus activity in the nasal cavity by activation of T-cell in Peyer's patch lymphocyte and stimulation of production of anti-influenza virus IgA antibody in nasal lymphocyte. When ovalbumin-sensitized allergic pulmonary inflammation model mice were administered orally with SST (1 g/kg) from 8 days before (11 times) or from 2 h after (4 times) to 4 days after the infection and infected with mouse-adapted influenza virus A/PR/8/34, replications of the virus in the both nasal and broncho-alveolar cavities or only nasal cavity were significantly inhibited at 5 days after infection in comparison with water-treated control by augmenting antiviral IgA antibody, respectively. These results suggest that SST is useful for both prophylaxis and treatment of influenza virus infection on patients with allergic pulmonary inflammation, such as bronchial asthma.     

Matching antibody class with pathogen type and portal of entry: cognate mechanisms regulate local isotype expression patterns in lymph nodes draining the respiratory tract of mice inoculated with respiratory viruses, according to virus replication competence and site of inoculation

Intranasal deposition of Sendai virus (SV) in C57BL/6 mice provokes an Ab-forming cell (AFC) reaction in mediastinal (MLN) and cervical lymph nodes (CLN), which drain the lungs and upper respiratory tract, respectively. While the majority of AFC elicited by infectious SV at both sites produced IgG, the CLN response to SV rendered inactive in replication was restricted almost entirely to IgA, although isotype switching in mediastinal continued to be skewed heavily to IgG. However, in vitro restimulation of the accompanying virus-specific T cell populations from the two sites did not reveal any significant difference in lymphokine output, and isotype expression was not altered substantially in mice lacking IL-4 or IL-6 genes. To dissociate the response to specific Ags from the inflammatory reaction to viral infection, we examined the response to inactivated SV in the face of infection with influenza virus A/HKx31. The magnitude and IgA dominance of the anti-SV AFC population in the CLN were unaffected by a simultaneous, vigorous, IgG-dominated CLN anti-influenza reaction. Evidently, the characteristics of this antiviral response are determined primarily by cognate interactions. Moreover, the IgA bias of the CLN AFC response to inactivated SV was observed only when the virus was delivered intranasally: injection under the epidermis of the cheek, a site that has a lymphatic drainage into the CLN, resulted in an IgG-dominated CLN AFC reaction, lacking IgA. The site of deposition of a vaccine can thus have more influence on the pattern of isotypes induced than the site at which the immune response is initiated.     

IgG2 associated hypergammaglobulinaemia in some Nigerians with HIV infection

Concentrations of immunoglobulins (IgA, IgG and IgM) were measured in Nigerians with (HIV) infection. Considerable elevations up to two-fold the reference values were observed for IgG and IgM in the patient group as a whole but elevations in IgA concentration were least marked albeit significantly different from the healthy subjects. Elevation of a particular isotype was not always concomitant with elevation of the other major classes in the same patient. Overall, these elevations were observed in both symptomatic and asymptomatic infected subjects. Further analysis of IgG hyperglobulinemia showed that increases in this major class may be due to increased IgG2 subclass concentrations. It is suggested that elevation of IgG2 subclass in Nigerians with HIV infection and not IgG1 or IgG3 may be due to genetic and environmental factors rather than variation in the strain of the virus.     

Influenza virus neutralizing antibodies and IgG isotype profiles after immunization of mice with influenza A subunit vaccine using various adjuvants

The influence of various adjuvants on the development of influenza virus neutralizing antibodies and distribution of anti-influenza virus IgG isotypes after immunization of mice with influenza A (H3N2) subunit vaccine was investigated. Serum titres of influenza virus neutralizing antibodies and titres of influenza specific IgG isotypes were determined by a neutralization enzyme immunoassay (N-EIA) and a cell-associated antigen enzyme immunoassay (CA-EIA), respectively. Serum antibody titres as measured by the two tests correlated highly (r = 0.82; P < 0.001). N-EIA titres were enhanced by 38- and 34-fold, when L180.5/RaLPS and FCA, respectively, were administered with 1 microgram of vaccine. The adjuvants Q-VAC, L180.5 [W/O/W], L180.5 alone and Montanide ISA 740 were only moderately or not effective in enhancing the immune response to the 1 microgram dose of vaccine. The Q-VAC and L180.5/RaLPS adjuvants favoured IgG2a and IgG2b isotype responses to influenza compared to the other adjuvants. We suggest that N-EIA and CA-EIA may be valuable tools to monitor the effects of adjuvants on the neutralizing antibody and antibody isotype responses after influenza vaccination.     

Trypanosoma cruzi infection in mice induces a polyisotypic hypergammaglobulinaemia and parasite-specific response involving high IgG2a concentrations and highly avid IgG1 antibodies

Trypanosoma cruzi infection in BALB/c mice induced a reversible polyisotypic hypergammaglobulinaemia, with particularly high levels of IgG2a, IgM and IgE. Hypergammaglobulinaemia started during the acute phase of infection and persisted during chronic disease until 11-13 weeks post-infection (w.p.i.), when immunoglobulin levels, with the exception of IgE, returned near normal values. Parasite-specific antibodies counted for 14 to 23% of gammaglobulinaemia, in acute and chronic infection respectively. The titres of IgM antibodies rose from two w.p.i. IgA, IgE and IgG subclass antibodies built up gradually over the time of parasite clearance (i.e., between three and six w.p.i.). All antibody isotypes, including IgM reached significant and stable titres throughout chronic infection. IgG2a, IgG1 and IgM antibodies had constantly higher titres than the other antibody isotypes. The dominance of IgG2a antibodies was due to their high plasma concentrations, around 70% of all antibodies available in the chronic infection. IgG1 had the highest functional avidity, whereas its concentration corresponded to only 10% of the whole antibody fraction. These results indicate that T. cruzi infection in mice induces a polyisotypic humoral immune response, dominated by some antibody isotypes, with major differences in concentrations and functional avidities. This could be of crucial importance in determining the outcome of infection.     

Analysis of anti-Trypanosoma cruzi antibody isotype specificities by western blot in sera from patients with different forms of Chagas' disease

Infection of humans with Trypanosoma cruzi leads to either a lifelong asymptomatic infection or to symptomatic presentations such as cardiomyopathy, mega-syndromes, or both. The reasons for the different clinical manifestations are not understood. We have previously studied a group of chronically infected individuals with different clinical forms of Chagas' disease and found that the levels of some anti-T. cruzi antibody isotypes, analyzed by enzyme-linked immunosorbent assay, differed among patients with different clinical presentations. We have expanded these studies to examine the antigen specificity of these patients' IgG1, 2, 3, IgM, and IgA by western blot. We observed that binding of particular antigens by some antibody isotypes were more prevalent in some clinical groups as compared to others. For example, IgG3 from 13 of 19 (68%) individuals with digestive manifestations bound a 68-kDa antigen, but only 3 of 31 (9%) individuals with cardiac involvement detected this same moiety. We also found that, regardless of the clinical group, the profiles of antigens recognized by each antibody isotype differs dramatically from the profiles recognized by each other isotype. Together with our previous observations demonstrating that the levels of anti-parasite antibody isotypes correlates with the clinical form, these data suggest that overall anti-T. cruzi antibody reactivities may indeed be skewed toward different antigens in individuals with different clinical presentations.     

Demonstration of immunoglobulin isotypes in the vitreous body as a contribution to the etiology of recurrent equine uveitis

The functional properties of different immunoglobulin isotypes in equine recurrent uveitis (ERU) has not been investigated yet. Here, we describe the quantitative determination of total immunoglobulin levels and isotype differentiation in the vitreous of four horses with ERU as compared to that of seven healthy horses. In contrast to almost equal amounts of total immunoglobulin in the vitreous of both groups, remarkable differences were found: All four of the horses with ERU had significantly higher IgA contents in their vitreous as compared to the control group. However, the other isotypes monitored (IgM, IgGa, IgGb, IgGc and IgG(T)) indicated no differences between both groups. Comparing the individual ratios of immunoglobulin isotypes in the vitreous and the autologous serum of two horses with ERU and two control animals provided informative results: IgM was only detected in serum but not at all in vitreous of all horses investigated. All four IgG isotypes monitored in the diseased animals as well as these IgG isotypes and the IgA in healthy animals were present in the same ratios in serum and in vitreous of the individual horses. In general, the content of such isotypes in the vitreous was about 1000 fold lower then the respective isotypes in the autologous serum. These results are compatible with a transfer of the IgG and IgA isotypes from the serum into the vitreous in healthy individuals. All four horses with ERU, however, had a selectively increased relative IgA content in the vitreous as compared to their serum. This argues for a preferential local IgA production within the framework of a local immunological reaction to antigens located within the eyes of horses with ERU.     

Interferon gamma-producing gammadelta T cell-dependent antibody isotype switching in the absence of germinal center formation during virus infection

Ig class switching usually occurs as a consequence of cognate interactions between antigen-specific B cells and CD4(+) alphabeta T cells. Vesicular stomatitis virus (VSV) infection of immunocompetent mice induces a rapid T-independent neutralizing IgM response followed by a long-lived T-dependent IgG response. Surprisingly, alphabeta T cell-deficient (TCRalpha-/-) mice also produced neutralizing IgG antibodies when infected with live VSV or with a recombinant vaccinia virus expressing the VSV glycoprotein (Vacc-IND-G), but not when immunized with UV-inactivated VSV (UV-VSV). The neutralizing IgG responses did not require the presence of NK cells or complement, but were crucially dependent on IFN-gamma and were predominantly of the IgG2a isotype. IgG production depended on residual CD3(+) non-alphabeta T cell populations present in the TCRalpha-/- mice, which produced IFN-gamma upon in vitro stimulation. A key role for gammadelta T cells was confirmed by the fact that TCRbeta-/- mice also generated strong neutralizing IgG responses to VSV, whereas TCRbeta-/-delta-/- mice produced very low titers. The neutralizing IgG responses of TCRalpha-/- mice were accompanied by the development of memory B cells, but not by antigen-specific germinal center (GC) formation. Thus, during viral infection of alphabeta T cell-deficient mice, gammadelta T cells may provide the signals that are required for isotype switching.     

Characteristics and responses of EBV immortalized B cells from periodontal disease patients

OBJECTIVE: This study was designed to examine human B cell responses to Actinobacillus actinomycetemcomitans (Aa). The general hypothesis to be tested was that Epstein-Barr virus (EBV) immortalized B cells could be used to investigate variations in B cell responsiveness of periodontitis patients to periodontal pathogens, and that B cells derived from the peripheral blood of periodontal disease patients infected with Aa demonstrate differences in in vitro activities compared to periodontally healthy subjects. DESIGN: EBV-transformed B cell lines were used to analyze immunoglobulin and Aa-specific antibody responses, as well as to determine the frequencies of cells producing immunoglobulin (Ig) of a specific isotype and detect clones secreting antibodies specific for Aa. Lymphoblastoid cells lines (LCL) were derived by clonal transformation of peripheral blood lymphocytes from 10 Aa-infected patients with adult periodontitis (Aa-AP) and seven normal subjects. METHODS: The B cells were incubated in Aa-coated polystyrene plates to separate adherent and non-adherent cells, and stimulate the cells with the whole bacteria. In addition, the B cells were stimulated with Aa LPS, E. coli LPS, or the polyclonal B cell activators (PBAs), pokeweed mitogen (PWM) and Staphylococcus aureus protein A (SpA). Both adherent and non-adherent cell populations were cultured for up to 15 days. MAIN OUTCOME MEASURE: Total immunoglobulins (Igs) and antibody (IgG, IgA, IgM) levels to Aa in the culture supernatants were assessed using an ELISA. The distribution of IgG, IgA, IgM and Aa-specific antibody producing cells was analyzed by a double immunoenzymatic staining technique. RESULTS: IgM levels produced by the LCLs were significantly increased vs IgG and IgA (P < 0.001). Three days after Aa stimulation, a marked increase in the level of total Igs and Aa-specific antibody was observed in adherent cells from Aa-AP (P < 0.05-0.03). Aa-specific antibody levels were significantly higher in the supernatants from Aa-AP vs normals throughout the culture interval (P < 0.03). There was also a significant increase in Aa-specific antibody levels after stimulation with Aa LPS or E. coli LPS (P < 0.05), whereas PWM and SpA had no significant effect on antibody to Aa. There was a predominance of IgM cells compared to IgG and IgA isotypes (P < 0.04) in LCLs from Aa-infected patients. After stimulation with Aa, a significant increase in the number of IgA (111%) and IgG (48%) secreting cells was observed, concomitant with a 74% decrease in the Ig-negative cell population. Total Aa+ cells increased significantly after stimulation (P < 0.001), predominated by Aa-specific IgG and IgM antibody producing cells. CONCLUSIONS: These results showed that LCLs from Aa-infected patients were polyclonal with respect to isotype distribution. Further stimulation with Aa revealed a shift to cytoplasmic IgG and IgA expression, as well as increases in the Aa-specific B cell population. In contrast, the PBAs stimulated the LCLs to synthesize primarily IgM. Additionally, the findings indicated that: (1) without T cells, polyclonal activation of B cells may lead to elevated Aa-specific B cell populations; and (2) the presence of previously sensitized B cells is required to exert an antigen specific antibody response in the LCL. We conclude that secondary activation of primed B cells by oral bacteria or their products in advanced periodontal lesions may contribute to the local accumulation of significant numbers of Ig-producing cells. This report also suggested that EBV-mediated transformation can be used to probe B cell-bacterial interactions in studies of periodontitis.     

Perceived benefits of and barriers to exercise and stage of exercise adoption in young adults

Antibodies to a wide spectrum of infectious agents belonging to the IgA, IgM and IgG isotypes are thought to be one of the protective factors in human milk. Cow milk-fed newborns are at an increased risk of infections as well as of allergic diseases and of necrotising enterocolitis. A reasonable approach would be to add to the milk formula fed to them the immunoglobulins present in human milk. We developed a pasteurised immunoglobulin preparation from pooled donor plasma ('Orabulin') containing 75% IgG, 18% IgA and 6% IgM for feeding to high-risk bottle-fed babies. Its molecular composition was studied by HPLC and by SDS-PAGE. The levels of IgA, IgG and IgM antibodies in Orabulin were compared to these in the immunoglobulin fraction of human colostrum and an enrichment was found. It is suggested that the presence of a standardised amount of human IgM in an immunoglobulin preparation intended for feeding to newborns may bring an additional advantage because of the high opsonising and virus-neutralising activity of the antibodies of this isotype.     

Role of immunoglobulin A monoclonal antibodies against P23 in controlling murine Cryptosporidium parvum infection

Cryptosporidium parvum is an important diarrhea-causing protozoan parasite of immunocompetent and immunocompromised hosts. Immunoglobulin A (IgA) has been implicated in resistance to mucosal infections with bacteria, viruses, and parasites, but little is known about the role of IgA in the control of C. parvum infection. We assessed the role of IgA during C. parvum infection in neonatal mice. IgA-secreting hybridomas were developed by using Peyer's patch lymphocytes from BALB/c mice which had been orally inoculated with viable C. parvum oocysts. Six monoclonal antibodies (MAbs) were selected for further study based on indirect immunofluorescence assay reactivity with sporozoite and merozoite pellicles and the antigen (Ag) deposited on glass substrate by gliding sporozoites. Each MAb was secreted in dimeric form and recognized a 23-kDa sporozoite Ag in Western immunoblots. The Ag recognized comigrated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis with P23, a previously defined neutralization-sensitive zoite pellicle Ag. MAbs were evaluated for prophylactic or therapeutic efficacy against C. parvum, singly and in combinations, in neonatal BALB/c mice. A combination of two MAbs given prophylactically prior to and 12 h following oocyst challenge reduced the number of intestinal parasites scored histologically by 21.1% compared to the numbers in mice given an isotype-matched control MAb (P < 0.01). Individual MAbs given therapeutically in nine doses over a 96-h period following oocyst challenge increased efficacy against C. parvum infection. Four MAbs given therapeutically each reduced intestinal infection 34.4 to 42.2% compared to isotype-matched control MAb-treated mice (P < 0.05). One MAb reduced infection 63.3 and 72. 7% in replicate experiments compared to isotype-matched control MAb-treated mice (P < 0.0001). We conclude that IgA MAbs directed to neutralization-sensitive P23 epitopes may have utility in passive immunization against murine C. parvum infection.     

Cellular mechanisms involved in protection against influenza virus infection in transgenic mice expressing a TCR receptor specific for class II hemagglutinin peptide in CD4+ and CD8+ T cells

Mice transgenic for a TCR that recognizes peptide110-120 of hemagglutinin of PR8 influenza virus in the context of MHC class II I-Ed molecules express the transgenes in both CD4+ and CD8+ T cells. We have found that these TCR-hemagglutinin (TCR-HA) transgenic mice display a significantly increased resistance to the primary infection with PR8 virus compared with the wild-type mice. The TCR-HA transgenic mice mounted significant MHC type II and enhanced MHC type I-restricted cytotoxicity as well as increased cytokine responses in both spleen and lungs after infection with PR8 virus. In contrast, the primary humoral response against PR8 virus was not significantly different from that of the wild-type mice. In vivo depletion and adoptive cell transfer experiments demonstrated that both CD4+ and CD8+ TCR-HA+ T cell subsets were required for the complete clearance of pulmonary virus following infection with a dose that is 100% lethal in wild-type mice. Whereas CD4+ TCR-HA+ T cells were necessary for effective activation and local recruitment of CD8+ T cells, CD8+ TCR-HA+ T cells showed a Th1-biased pattern and MHC type II-restricted cytotoxicity. However, in the absence of in vivo expression of MHC type I molecules on the infected cells, the protection conferred by the TCR-HA+ T cells was impaired, indicating that the enhanced MHC class I-restricted cytotoxicity due to TCR-HA+ CD4+ Th cells was a critical element for clearance of the pulmonary virus by the transgenic mice.     

Culture and characterization of equine terminal arch endothelial cells and hoof keratinocytes

The occurrence of abnormally low serum immunoglobulin (Ig) levels is well-known in B chronic lymphocytic leukemia (CLL), but published data on IgG subclass levels are virtually absent. We measured serum IgG subclass levels in 52 B CLL outpatients, most in stage A and untreated, using an indirect immunoenzymatic assay with monoclonal antibodies. Mean levels of all Ig isotypes were lower than in normal controls in the whole group of patients, except for IgG2 in those studied at diagnosis. Levels of IgG1, IgG2, IgA, and IgM were lower in patients with a long disease duration than in those studied earlier. IgG subclass deficiencies occurred in 54% of cases and the most frequently affected isotype was IgG1. Every possible combination of IgG subclass and Ig class deficiencies from the selective deficiency of a single subclass to a combined deficiency of all isotypes was observed. This marked heterogeneity argues against the occurrence of isolated defects of one of the cytokines involved in Ig switching as a cause of hypoimmunoglobulinemia in CLL.     

Monoclonal and polyclonal antibodies to chicken immunoglobulin isotypes specifically detect turkey immunoglobulin isotypes

Turkey immunoglobulin (Ig) isotypes IgG and IgM were isolated from blood and IgA was isolated from bile. Isolation was accomplished by gel filtration of the ammonium sulphate cut on Sephacryl S-200. Using immunoelectrophoresis and indirect ELISA, the cross-reactivity between antibodies, of monoclonal and polyclonal origin, specific for the Ig isotypes of chicken, and the purified turkey Ig isotypes was evaluated. Commercially available polyclonal antibodies, anti-chicken/IgA (alpha-chain specific, affinity purified), anti-chicken/IgG (Fc-fragment specific) and anti-chicken/IgM (mu-chain specific) showed an interspecies cross-reactivity with the corresponding turkey Ig isotypes. The monoclonal antibody (MAb) AV-G3 specifically detected turkey IgG, whereas MAb M1 reacted exclusively with turkey IgM. This panel of anti-immunoglobulins represents a useful tool for examining the humoral immune responses of turkeys.     

The pathogenesis of alcoholic liver disease

The immunoglobulin (Ig) isotype (IgG, IgA, IgM) production of peripheral blood mononuclear cells from 28 patients having multiple myeloma (MM) was analyzed. The total Ig secreting capacity of the cells, as measured by ELISA from the cell culture medium, was not found to be significantly reduced in MM (1,118 +/- 1,394 micrograms/l) as compared to the values of 9 controls (898 +/- 520 micrograms/l), but a significant isotype switching towards the tumor paraprotein type was observed in the patients with active MM (p < 0.001). The percentage of IgG in the active IgG-MM was 88 +/- 11% and that of IgA in the active IgA-MM 83 +/- 13%, the control values being 44 +/- 11% for IgG and 44 +/- 13% for IgA. The proportions of isotypes resembled those of the controls in the inactive phase of the disease. Despite this dominating paraprotein class isotype production, no evidence of Ig gene clonal rearrangements was found in cells studied by either Southern blotting or the more sensitive polymerase chain reaction method, which suggests that polyclonal rather than monoclonal PB B cells are responsible for the Ig production observed.