Biomedical subjects

C B Reimer

Publications and source records attributed to C B Reimer.

At least 19 recordsLinked to original sources

PubMed1996

Evaluation of monoclonal antibodies with specificity for human IgA, IgA subclasses and allotypes and secretory component. Results of an IUIS/WHO collaborative study.

51 monoclonal antibodies (McAb) with putative specificity for human IgA, the IgA subclasses, Am allotypes or secretory component (SC) were evaluated for immunoreactivity and specificity by nine laboratories employing immunodiffusion, agglutination, immunohistological assays, immunoblotting and direct binding and competitive inhibition enzyme immunoassays. McAbs specific for IgA PAN (n = 24), IgA1 (n = 7), IgA2 (n = 3), IgA2m(2) (n = 2), non-IgA2m(2) (n = 4) and SC or secretory IgA (n = 5) were identified that were immunoreactive and specific in the assays employed. The McAbs identified as IgA- or SC-reactive were shown to be non-reactive to human IgG, IgM, IgD, IgE, kappa and lambda by direct binding and competitive inhibition immunoassays. Interestingly, no McAbs with restricted specificity for IgA2m(1) were identified. Some McAbs displayed higher affinity and/or better performance in one or several of the assay groups. The IgA- and SC-specific McAbs identified in this international collaborative study have potential as immunochemical reference reagents to identify and quantitate monomeric and polymeric IgA in human serum and secretions.

Animals↗

PubMed1993

Cross-reactivity of 75 monoclonal antibodies to human immunoglobulin with sera of non-human primates.

We systematically analyzed a panel of 75 murine monoclonal antibodies (mAbs) reactive with human immunoglobulins IgG, IgA, IgM, IgD, and kappa and lambda light chains for reactivity with serum immunoglobulins of higher primates. In the great apes, and to a lesser extent in other primates, epitopes related to human light chains, IgM, IgA, IgD, and all 4 IgG subclasses were identified with many of the mAbs. Those mAbs identified as reactive with a given species may be useful for immunologic studies of these species. Cladistic analysis of antigenic relatedness generated a phylogenetic tree consistent with current anatomic or molecular taxonomies.

Animals↗

PubMed1992

Evaluation of monoclonal antibodies having specificity for human IgG subclasses: results of the 2nd IUIS/WHO collaborative study.

Following the 1st IUIS/WHO Collaborative Study of monoclonal anti-IgG subclass antibodies, a panel of WHO Specificity Reference Reagents (SRR) was established [Jefferis, R., et al. (1985) Immunol. Lett., 10, 223]. At the time, the hope was expressed that further reagents particularly for IgG2, and other allotypic specificities would become available which could be applied in a wide range of assay protocols. The 2nd study reports the evaluation of nineteen anti-subclass and seven anti-allotype monoclonal antibodies. The anti-IgG1 antibody HP6187 was equivalent in performance to the SRR. Others, that were not of the mouse IgG1 isotype, may be useful for particular applications. The anti-IgG2 antibody HP6200 could be a valuable addition to the WHO SRR; it is specific for an epitope in the Fab region but does not have the light chain bias of HP6014. Antibodies of putative allotype specificity exhibited the claimed specificity when used within protocols similar to those employed by the originating laboratory. It appears to be inherent in the nature of the epitopes (allotopes) recognized that it will take several years before reagents applicable to a wide range of techniques will become available.

Antibodies, Anti-Idiotypic↗

PubMed1992

Immunoglobulin class and subclass-specific monoclonal antibody sandwich ELISA for the detection of antibodies against coxsackieviruses B, types 1-5.

Immunoglobulin subclass-specific ELISAs were developed for human IgG1, IgG2, IgG3, IgG4, IgAtotal, and IgM directed against Coxsackie B (CB) virus types 1, 2, 3, 4, and 5. In all the assays the solid phase was coated with immunoglobulin class/subclass-specific monoclonal antibodies, followed by an incubation with the serum specimens. Incubation with one of the CB viruses, as well as an incubation with biotinylated serotype-specific monoclonal antibodies to the same virus type provided the virus specificity. Finally, there were incubations with peroxidase labeled Extravidin and the substrate-chromogen system. This ELISA method eliminated the competition between the immunoglobulin classes and subclasses. IgG3 and/or IgG1 were seen most frequently of the IgG subclasses, but IgG2 and IgG4 were also present infrequently. The viral specificity of the antibody subclass assays seems to be predominantly at the enterovirus group level, but this remains to be evaluated in a larger study. IgA and IgM were seen almost exclusively in specimens from patients with acute enteroviral infections, except in the assays with the crude CB5 antigen. This indicates the possible suitability of the IgA and IgM assays as diagnostic tests for enteroviral infections. A larger study is necessary to confirm this finding.

Adult↗

PubMed1991

Guatemalan human onchocerciasis. I. Systematic analysis of patient populations, nodular antigens, and specific isotypic reactions.

The population from five Guatemalan plantations in areas endemic for onchocerciasis was surveyed, and 1032 individuals were recruited to participate in our study. From physical examination, past clinical history (5 to 8 yr), laboratory evidence and sample availability, a group of 778 long term residents with confirmed disease status were selected for detailed examination. We were able to identify 268 long term residents of endemic areas who had never been infected, 44 of these are from hyper- and mesoendemic areas. The 44 uninfected individuals from the hyper- and mesoendemic areas, because of their considerable exposure to this disease, were classified as "putatively immune." Intact nodules containing adult worms of Onchocerca volvulus were homogenized in the presence of protease inhibitors and fractionated into particulate and aqueous isotonic soluble antigens. Systematic analysis of these Ag fractions showed considerable amounts of Ig, presumably associated with Ag in the form of immune complexes. Individual specific antibody reactions from all 778 patients to nodule Ag were examined. Reactions to O. volvulus antigens by antibodies from patients with confirmed parasitic infections were almost exclusively restricted to IgG1 and IgG4 isotypes. Antigenic activity appeared to be primarily associated with low molecular mass (14 to 29 kDa) components. Some competitive blocking of antibody activities of other isotypes by IgG1 was observed, most notable was that of IgG3 and IgA. IgG4 and IgM activities were not significantly blocked.

Adolescent↗

PubMed1991

Guatemalan human onchocerciasis. II. Evidence for IgG3 involvement in acquired immunity to Onchocerca volvulus and identification of possible immune-associated antigens.

Ag-specific isotypic differences in immune response to Onchocerca volvulus Ag were assessed for 778 long term residents of endemic Guatemalan areas by quantitative ELISA with 5-min incubation steps and immunoblot. The study population was separated into five groups based on clinical status: N+F+, N+F-, N-F+, N-F-H+, and N-F-H-, where N = O. volvulus adults (nodule), F = microfiladermia, and H = history of O. volvulus infection. A subset of 44 individuals with high exposure to onchocerciasis from the N-F-H- group were critically evaluated and designated as "putatively immune." IgG1 reactivity to O. volvulus Ag was elevated in the majority of infected persons, but not in putatively immune individuals. Specific IgG3 levels, however, were equally elevated in all groups. The majority of N+F- persons also had elevated IgG1 levels, but they were lower than those found in F+ persons. IgG3 reactivities to a group of antigens at 20 kDa (GP20) were seen in many uninfected persons and some N+F- persons. In contrast, most F+ persons, react to this Ag with IgG1 and not IgG3. A mangabey inoculated with the infectious larval stage of O. volvulus (L3), but showed no signs of infection, began to recognize GP20 at 2 wk postinoculation. Early recognition of GP20 was possibly elicited by the larval stage. Purified nodule Ag from N+F+ individuals contained GP20, however, identical nodule Ag prepared from N+F- individuals did not. These data suggest that GP20 Ag may be common to both uterine microfilaria and the infectious larval stages. The fact that GP20 is predominantly recognized by IgG3 in putatively immune persons and some N+F- persons suggests that this increased IgG3 activity may be important in acquired immunity to onchocerciasis.

Adolescent↗

PubMed1991

Immunoglobulin G subclass values in healthy black and white children.

To determine whether IgG subclass concentrations differed between healthy black and white children, we measured IgG1, IgG2, IgG3, and IgG4 immunoglobulins by enzyme-linked immunosorbent assay in sera from 246 black children aged 6 to 42 months. We then compared these values with the normal values established for 664 white children aged 6 to 60 months. The IgG1, IgG2, and IgG4 subclass concentrations of the black children were lower than those for white children; many of the values were below the 95% confidence limits established for white children: 46 (19%) of 246 IgG2 values and 19 (8%) of 246 IgG4 values for black children were below the normal limits. We compared the geometric mean values for black and white children, as determined for each 6-month age grouping between 6 and 42 months of age; 367 of the 664 white children were less than 42 months of age and were included in this analysis. The geometric mean values for IgG1, IgG2, and IgG4 levels were consistently lower for black children than for white children. The differences were significant for IgG1 subclass values of those children older than 24 months and for IgG2 and IgG4 values of those children older than 18 months. No consistent differences were noted for IgG3 subclass values. We conclude that young black children have lower IgG1, IgG2, and IgG4 serum concentrations than are found in white children. If normal IgG values for white children are used, healthy black children may be erroneously classified as IgG subclass deficient. The mechanism and biologic relevance of these population differences need to be evaluated.

Black People↗

PubMed1991

Human markers for IgG2 and IgG4 appear to be on the same molecule in the chimpanzee.

It has been reported that all four immunoglobulin G (IgG) subclasses present in human serum are also present in chimpanzee serum, as detected with antibodies specific for the human IgG subclasses. We used monoclonal antibodies (mAb) specific for human IgG subclasses to measure concentrations of the four subclasses in the chimpanzee sera. Initial ELISA studies indicated that epitopes for all four human subclasses are present in chimpanzee sera. The concentrations of IgG1, IgG2 and IgG3 were similar in human and chimpanzee sera, but the registered concentrations of IgG4 were different. Absorption of IgG2-reactive material from chimpanzee serum with IgG2 mAb resulted in removal of IgG4-reactive material as well. Conversely, absorption of IgG4-reactive material removed IgG2-reactive material. IgG2-reactive material, isolated from chimpanzee serum using solid-phase anti-IgG2 mAb, reacted with anti-IgG4 mAb, and isolated IgG4-reactive material reacted with anti-IgG2 mAb. Three anti-IgG2 mAb and five anti-IgG4 mAb, each of which react with separate epitopes on their respective human isotype, were used in these studies. We conclude that chimpanzee serum contains only three IgG isotypes related to those of humans, one of which contains determinants related to both human IgG2 and IgG4.

Adult↗

PubMed1990

Interpretation of IgG subclass values: a comparison of two assays.

Because we have noted discordant results in the measurement of IgG subclass concentrations by means of a widely available commercial radial immunodiffusion (RID) kit in comparison with an enzyme-linked immunosorbent assay (ELISA) developed at the Centers for Disease Control (CDC), we conducted in a blinded manner a comparison of the two assays, using sera from 48 healthy children. The correlation coefficients between the assays were 0.92, 0.82, 0.93, and 0.86 for the IgG1, IgG2, IgG3, and IgG4 assays, respectively. However, the RID assay assigned lower values for IgG1 and IgG4 determinations than the ELISA did. Furthermore, the "normal lower range values" provided by the RID assay were higher for each IgG subclass. When the sera from the healthy control subjects were analyzed with the RID assay, 12 (25%) of 48 subjects had values below the normal range for at least one subclass measurement. In contrast, with the CDC ELISA, all values were within the 95% confidence limits determined for the CDC ELISA. We suggest that age-specific normal limits be established with the use of sera from many healthy subjects for any assay measuring IgG subclass concentrations. As new groups of immunodeficiencies are defined and potential therapies are advocated, careful attention to assay standardization will result in a clearer delineation of these disease groups and of their response to treatment.

Child↗

PubMed1990

IgG subclasses in the serum and skin in subacute cutaneous lupus erythematosus and neonatal lupus erythematosus.

IgG subclasses differ in their biologic and chemical properties, such as complement fixation, protein and cellular binding, and placental transfer. In this study, IgG subclasses of anti-Ro/SSA antibodies in subacute cutaneous lupus (SCLE) and neonatal lupus (NLE) are examined in the serum and in the skin. IgG subclasses in NLE beginning in utero (NLE-heart disease) are compared to subclasses in NLE beginning after birth (NLE-skin disease). Human skin was grafted onto athymic mice, mice were injected with one of eight anti-Ro/SSA maternal NLE sera (four heart block, four skin disease) or seven anti-Ro/SSA SCLE sera, and grafts were examined for IgG subclasses using monoclonal anti-human IgG subclass reagents in an immunofluorescent technique. Lesional skin was examined from four SCLE patients. IgG1 was the only IgG subclass detected in the grafts and skin lesions. IgG1 was the predominant anti-Ro/SSA IgG subclass detected in SCLE and NLE sera in an ELISA using a synthetic Ro/SSA polypeptide. These studies show that the maternal anti-Ro/SSA autoantibodies in NLE-heart disease sera are predominantly IgG1 and are therefore likely to be present in the fetus at the time of gestation, when heart block usually develops. Second, differences in the clinical presentations of NLE (in utero vs. postnatal disease) cannot be attributed to differences in anti-Ro/SSA IgG subclasses. Finally, the subclass bound in the skin in SCLE is IgG1, a subclass capable of mediating tissue injury via complement or cellular effectors.

Animals↗

PubMed1990

IgG heavy-chain subclass restriction of thyrotropin-binding inhibitory immunoglobulins in Graves' disease.

The IgG subclass composition of antibodies is an important determinant of their function. Thyrotropin receptor antibodies cause the hyperthyroidism of Graves' disease but their subclass distribution has been incompletely investigated. We have therefore purified IgG subclasses from Graves' sera by passage over affinity columns designed to deplete all but a single subclass, and then assayed those pure subclass fractions for their ability to displace radiolabelled thyrotropin from its solubilized receptor as a measure of thyrotropin receptor antibody activity. Sufficient activity was recovered for analysis in nine of 10 Graves' patients, in five of whom activity was almost completely (97-100%) restricted to the IgG1 subclass; in the remaining four patients the response was predominantly IgG1 and IgG4 with marked under-representation of the IgG2 subclass. This contrasts with the unrestricted subclass response, in the same fractions, for autoantibodies against thyroglobulin and microsomes. These results suggest that there may be a primary defect at the B-cell level in Graves' disease.

Autoantibodies↗

PubMed1990

Thyroid-stimulating antibody activity between different immunoglobulin G subclasses.

To investigate the distribution of thyroid-stimulating antibody (TSAb) activity between IgG subclasses, sera from 11 patients with Graves disease (including the National Institute of Biological Standards and Control (NIBSC) Research Standard, long acting thyroid stimulator-B) were fractionated by chromatography on affinity columns of monoclonal IgG subclass antibodies or protein A to deplete all but a single subclass. The resulting fractions were 98% or more pure for a single subclass. In all 11 patients, TSAb activity appeared to be confined to the IgG1 fraction as determined by cAMP production on addition of the fractions to the FRTL-5 rat thyroid cell line. In all of eight specimens from seven patients so tested, the whole serum activity was recovered in the IgG1 fraction, after adjusting for the recovery of the isotype from the column. TSAb activity in one serum comprised both lambda and kappa light chains but was IgG1 restricted. This IgG subclass restriction was not found when the same fractions were tested for thyroglobulin, microsomal/thyroid peroxidase, or tetanus toxoid antibody activity. Together with previous results showing marked restriction of both light chain usage and isoelectric point of TSAb, these results support the idea that Graves' disease may be the result of an oligo- or possibly monoclonal response at the B cell level.

Cyclic AMP↗

PubMed1989

Specificity and association constants of 33 monoclonal antibodies to human IgA epitopes.

We used an immunofluorescent sequential-saturation-of-antibody assay and an interactive computer program for Scatchard analysis to determine association constants (Ka) of 33 murine monoclonal antibodies (Mabs) specific for human IgA epitopes. Ka ranged from 0.37 to 690 x 10(7) liters per mole (an approximate 1900-fold difference). Specificity was validated with a panel of 18 highly purified IgA1 and IgA2 myeloma proteins and secretory IgA using an immunofluorometric assay. Western blots of bacterial IgA protease digests were used to locate the epitopes of IgA specific Mabs in either the Fab, Fc, or hinge region. Mabs specific for unique epitopes on secretory IgA or free secretory component (FSC) were produced and evaluated.

Animals↗

PubMed1989

Immunoassay for serologic diagnosis of influenza type A using recombinant DNA produced nucleoprotein antigen and monoclonal antibody to human IgG.

Influenza type A nucleoprotein (NP) derived from the full length cloned gene expressed in E. coli was evaluated in a solid phase enzyme immunoassay (EIA) for detection of human antibody to influenza. Monoclonal antibody to human IgG was used for detection. Direct and indirect assays were developed and sera were tested in serial and single dilution formats. Preliminary results indicated that recombinant-and virion-derived NP antigens were comparable in binding ability to plastic and binding human antibody. Eighty-seven paired sera from influenza patients were tested. The most sensitive assay (indirect-serial dilution) detected 56 (64%) rises and the simplest assay (direct-single dilution) detected 43 (49%) rises, compared to 36 (41%) for complement fixation. Paired sera from 18 control patients showed no evidence of antibody rises by any of the assays. Forty-nine paired sera from influenza B infected patients were negative for antibody rises except for one borderline rise by the indirect-serial dilution assay. These results indicate that the use of recombinant DNA derived nucleoprotein for immunoassay is feasible. The sensitivity of immunoassays using NP adsorbed to the solid phase and monoclonal antibody specific for human IgG to detect bound antibody exceeded that of conventional complement fixation testing for establishing serologic evidence of influenza type A infection.

Antibodies, Monoclonal↗

PubMed1989

Human antibody responses to bacteriophage phi X 174: sequential induction of IgM and IgG subclass antibody.

We studied the appearance of antigen-specific immunoglobulin classes and IgG subclasses in normal adult human subjects in response to primary, secondary, and tertiary immunization with the T-cell-dependent neo-antigen bacteriophage phi X 174. To complete the study we developed a sensitive, specific, and reproducible ELISA assay which was closely comparable to the widely used neutralization assay for total antibody (r = +0.97) and for IgG antibody (r = +0.93), and reasonably comparable for IgM antibody (r = +0.76). We confirmed that the initial response to primary immunization was predominantly, but not exclusively, IgM antibody. The secondary and tertiary responses demonstrated memory, amplification, and switch from IgM to IgG antibody. There was an orderly appearance of phage-specific IgG subclasses. IgG3 and IgG1 antibodies appeared 2 to 6 weeks after primary immunization. In all subjects there was a marked increase in IgG1 and IgG3 antibody after secondary immunization, and IgG2 antibody followed closely; IgG4 antibody appeared in some subjects. IgM antibody persisted in significant amounts (approx 50%) throughout the secondary response period. Following tertiary immunization, IgG1, IgG2, and IgG3 antibody consistently increased, and IgG4 antibody appeared in all subjects; IgG1 antibody predominated. Low levels of IgM antibody (approx 1% of total) persisted during the tertiary response. The persisting antibody on long-term follow-up (median 4 years after immunization) was virtually all (greater than 90%) IgG1.

Adult↗

PubMed1989

Affinity purification of IgG subclasses and the distribution of thyroid auto-antibody reactivity in Hashimoto's thyroiditis.

To delineate accurately the IgG subclass distribution of thyroid auto-antibodies, sera from nine patients with Hashimoto's thyroiditis were fractionated into IgG subclasses by complete depletion of the other IgG subclasses on affinity columns. All IgG subclass fractions contained thyroglobulin and microsomal (or thyroid peroxidase) antibody activity, although when compared to the total serum concentrations of IgG subclasses, IgG4 antibodies were overrepresented. However, in contrast to recent studies, this particular subclass never predominated--IgG4 antibody levels being exceeded by those of the IgG1 and IgG2 subclasses; it seems likely that these differences relate to varying sensitivity for different subclasses in previously used assay methods. This pattern of subclass activity differed from that of tetanus toxoid antibodies, which were found in six subjects. There was no light chain restriction within any subclass, showing that the overproduction of IgG4 thyroid antibodies is not of monoclonal origin. The functional affinity of subclasses for both thyroid antigens varied between patients, but IgG2 subclass fractions showed the highest functional affinity in the majority of samples. We also found that IgG2 subclass thyroid antibodies were ineffective in eliciting antibody-dependent cell-mediated cytotoxicity, as distinct from the other three subclasses. Our results show that thyroid antibodies are less restricted in their IgG subclass distribution and patients are less heterogeneous than previously described. Moreover, IgG2 thyroid antibodies are quantitatively important and differ in relative functional affinity and effector function from IgG1 and IgG4 thyroid antibodies.

Adult↗

PubMed1989

Serum immunoglobulin G antibody subclass response to respiratory syncytial virus F and G glycoproteins after first, second, and third infections.

Serum samples from 31 children who experienced two or three infections with respiratory syncytial virus (RSV) in the first four years of life were tested in an enzyme-linked immunosorbent assay to examine the immunoglobulin G (IgG) subclass responses to the RSV F and G surface glycoproteins associated with primary infection and reinfection. We sought to determine whether the greater degree of glycosylation of the G glycoprotein was reflected in an IgG subclass immune response more like that to a polysaccharide antigen than to a protein antigen. We found that the IgG1/IgG2 ratio of postinfection antibody titers to F was fourfold higher than that to the G glycoprotein after RSV infections 1, 2, and 3. The IgG2 response to the heavily glycosylated G glycoprotein differed from that to a polysaccharide antigen in that the IgG1/IgG2 ratio remained constant with age, whereas the response to a polysaccharide antigen decreased as the IgG2 response increased with age. We also noted that antibody responses to both surface glycoproteins in the IgG1 and IgG2 subclasses reached their maximum levels after RSV infection 2.

Antibodies, Viral↗

PubMed1988

Two-site immunoenzymometric assays for serum IgG subclass infant/maternal ratios at full-term.

Improvements were made in the range, precision, convenience, automation, and reliability of our previously published two-site immunoenzymometric assays using mouse monoclonal antibodies specific for human IgG and its subclasses. The serum concentration ratios for these immunoglobulin isotypes were measured in neonatal/maternal paired sera from 119 normal full-term deliveries. These ratios are significantly different than 1.0 (P = 0.001) for total IgG, IgG1, and IgG2 (show non-equality of paired neonatal/maternal sera concentrations) but are not significantly different than 1.0 for IgG3 and IgG4. On average, IgG1 is elevated 61%, and IgG2 is depressed 11% in the full-term neonate with respect to its own mother. In some pregnancies, active transport of IgG1 may be selectively enhanced by low material IgG1 concentration and selectively inhibited by high levels.

Antibodies, Monoclonal↗