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C B Reimer

Publications and source records attributed to C B Reimer.

At least 37 records · Page 2Linked to original sources
PubMed1988

Estimation of association constants of 42 monoclonal antibodies to human IgG epitopes using a fluorescent sequential-saturation assay.

We measured the association constant (Ka) for 42 murine monoclonal antibodies (MAbs) to human IgG epitopes. Included are antibodies, previously evaluated in an IUIS/WHO collaborative study, to various epitopes on the four subclasses of human IgG - IgG Fc, IgG Fab, kappa, and lambda - and to selected IgG allotopes. We used a sequential-saturation immunofluorescent assay and interactive computer program to determine the Ka by Scatchard analysis. Kas ranged from unmeasurably low by this method (approximately 10(6) L/M) to 3.8 X 10(9) L/M. Some class specific MAbs had large Ka differences for different subclasses and some subclass specific MAbs had large Ka differences for molecules of the same subclass but of different light-chain types.

Antibodies, Monoclonal↗
PubMed1988

Clinical and immunologic responses to the capsular polysaccharide of Haemophilus influenzae type b alone or conjugated to tetanus toxoid in 18- to 23-month-old children.

The safety and immunogenicity of Haemophilus influenzae type b (Hib) capsular polysaccharide (CPS) alone, or covalently bound to tetanus toxoid in saline solution (Hib-TT) or adsorbed onto AI(OH)3 (Hib-TT ads), were evaluated after one injection into 18- to 23-month-old healthy children in Sweden. No side reactions were elicited by Hib CPS; side reactions elicited by the two conjugates were similar and comparable to those reported for diphtheria and tetanus toxoids adsorbed. Hib-TT was the most immunogenic of the three vaccines, eliciting about 10-fold higher antibody levels than Hib CPS; of 28 vaccinees, all had greater than 1.0 microgram Ab/mL serum after immunization with Hib-TT. Increases of Hib CPS antibodies within immunoglobulin classes induced by the three vaccines were, in decreasing order, IgG greater than IgM greater than IgA. Within IgG subclasses, rises in IgG1 Hib CPS antibodies were the most frequent, followed by IgG2; some vaccinees with high postimmunization levels also had rises in IgG3 and one in IgG4. Immunization-induced Hib CPS antibodies were bactericidal. Hib-TT also elicited higher levels of tetanus toxoid antibodies than Hib-TT ads; these tetanus toxoid antibodies neutralized tetanus toxin in vivo.

Aluminum Hydroxide↗
PubMed1988

Immunoglobulin class and subclass profile of the Ro/SS-A autoantibody response.

We have developed an enzyme-linked immunosorbent assay (ELISA) for autoantibody to Ro/SS-A antigen (anti-Ro/SS-A) in order to more fully characterize the autoimmune response that occurs to this antigen in patients with subacute cutaneous lupus erythematosus (SCLE). The microtiter plate-immobilized, biochemically purified Ro/SS-A antigen reacted with anti-Ro/SS-A antibody, but not with other closely related specificities (anti-La/SS-B, anti-SM, anti-U1-RNP) or normal sera. The optimal pH of antigen-antibody reaction in this ELISA was 7.2. The binding of sera containing anti-Ro/SS-A was inhibited 80% by preincubation with the same amount of Ro/SS-A antigen used for coating the plate. Although 11 of the 14 (79%) SCLE sera studied had precipitating anti-Ro/SS-A antibody by immunodiffusion, 13 (93%) sera had abnormally elevated IgG, IgA, or IgM ELISA binding levels. A good correlation between IgG anti-Ro/SS-A ELISA binding levels and immunodiffusion titers was observed (r - 0.8588, p less than or equal to 0.001) suggesting that IgG is the major anti-Ro/SS-A antibody class detected by double immunodiffusion, Sera with a combination of high rheumatoid factor levels (latex 3+ or higher) and high anti-Ro/SS-A titers (1:8 or higher in immunodiffusion) tended to give an abnormally high IgM anti-Ro/SS-A ELISA binding levels. After rheumatoid factor activity was removed by absorption with heat-aggregated human IgG, a 50% decrease in IgM anti-Ro/SS-A ELISA binding was noted. On the other hand, absorption of rheumatoid factor-negative sera that contained high IgM anti-Ro/SS-A binding activity did not significantly decrease ELISA binding levels. Prednisone and 6-azathioprine reduced the level of IgG anti-Ro/SS-A autoantibody in sera of treated SCLE patients by 50%. The IgG subclass profile of anti-Ro/SS-A autoantibody was analyzed by using mouse monoclonal antibodies specific for the 4 human IgG subclasses. Of anti-Ro/SS-A positive SCLE sera, 91% had predominantly IgG1 subclass autoantibody. The coexistence of IgM and IgG anti-Ro/SS-A autoantibody and the predominance of the IgG1 subclass is compatible with the possibility that this autoantibody response is under T-cell control. The predominance of IgG1 in the autoimmune response to Ro/SS-A antigen in SCLE patients is consistent with the hypothesis that antibody dependent cell mediated cytotoxicity could be an important immunologic effector mechanism in this disorder.

Animals↗
PubMed1988

Immunoblot analysis of humoral immune responses following infection with Bordetella pertussis or immunization with diphtheria-tetanus-pertussis vaccine.

To help develop better diagnostic tests for pertussis, we examined the serologic response to whole-cell proteins of Bordetella pertussis after natural infection or vaccination with diphtheria-tetanus-pertussis vaccine. Serum specimens collected during a pertussis outbreak investigation and from uninfected persons were used in Western blot (immunoblot) analyses to determine the presence of immunoglobulin G (IgG) and IgA antibodies to specific B. pertussis proteins. IgG antibodies to proteins of molecular masses 220 and 210 kilodaltons (kDa) were detected in 14 of 18 serum samples obtained from patients with culture-confirmed pertussis greater than or equal to 40 days after the onset of coughing. IgA antibodies were detected in 15 of the 18 samples. Of 19 serum samples obtained from patients who had not been ill with pertussis, 6 contained IgG antibodies to these proteins and 1 contained IgA antibodies. The two proteins bound antiserum specific for filamentous hemagglutinin and comigrated with purified filamentous hemagglutinin. IgG antibodies to two additional protein bands of molecular masses 84 and 75 kDa were associated with previous vaccination. Antibody to the 84-kDa protein was detected in 15 of 17 vaccinated, never-infected persons, and antibody to the 75-kDa protein was detected in 16 of the 17. None of 11 nonvaccinated, never-infected persons tested had antibodies to either protein. All seven fully vaccinated persons with culture-documented infection had antibodies to both proteins. Antibodies to the 84-kDa protein were detected in 6 of 22 nonvaccinated and infected persons, and antibodies to the 75-kDa protein were detected in 8 of the 22. Use of Western blot analysis in this study allowed us to distinguish antibody responses to infection and immunization.

Adolescent↗
PubMed1988

Hypergammaglobulinemia associated with human immunodeficiency virus infection.

The serum concentrations of 11 Ig isotypes (IgG, IgG1, IgG2, IgG3, IgG4, IgA, IgA1, IgA2, IgM, IgD, and IgE) were measured in four relatively small groups of homosexual (or bisexual) males. All these patients were seropositive for HIV. Two of the groups (nonprogressors) were clinically stable for approximately 2 years and were characterized either as asymptomatic or with PGL. The third group (progressors) developed AIDS 2-38 months after blood specimens were taken. The fourth group had AIDS. A fifth group of anti-HIV-seronegative heterosexual males completed the study. The geometric mean IgA serum concentration was more markedly elevated over normal control sera than any of the other study groups and was the only Ig isotype that was significantly higher in the progressor than in the nonprogressor group. The geometric IgG1 serum concentration was significantly higher in asymptomatic nonprogressors, PGL-nonprogressors, progressors, and AIDS patient groups than that in HIV-seronegative normals. In contrast, the geometric mean IgG2 serum concentration is depressed in all the anti-HIV-seropositive patients (but not significantly with the AIDS group). Multivariate analysis showed the Ig-isotype assays to have much less predictive power for progression to AIDS than the T-helper cell assays.

Acquired Immunodeficiency Syndrome↗
PubMed1987

Quality control of murine monoclonal antibodies using isoelectric focusing affinity immunoblot analysis.

Quality control of murine hybridoma secretory products was performed using two variations of the isoelectric focusing affinity immunoblot analysis. The first approach employed antigen-coated nitrocellulose placed on top of an acrylamide gel containing isoelectrically focused ascites to bind antigen specific monoclonal antibody (MoAb). Murine antibody bound to the insolubilized antigen was then detected with enzyme-conjugated anti-mouse IgG. In a second variation, focused ascites proteins were passively blotted onto nitro-cellulose and specific monoclonal antibody was detected with enzyme-conjugated antigen. Several batches of ascites containing anti-human IgG antibodies that were produced by 6 hybridomas over a 1-3 year period were assessed by IEF-affinity immunoblot analysis. Both immunoblot approaches permitted effective monitoring of immunoreactive antibody for pI microheterogeneity. IEF-affinity immunoblot patterns of unprocessed ascites displayed specific MoAb banding patterns with narrow pI ranges (less than or equal to 0.6 pH units), in contrast to the reported 5.5-8.0 pI range of polyclonal mouse IgG. Banding patterns obtained in the IEF affinity immunoblot typically displayed 3-5 major dense bands flanked by 2-4 minor fainter bands. Batches of ascites obtained years apart produced similar immunoblot patterns, indicating constant antibody production and confirming the stability of these hybridoma clones. Minor bands appeared in 2 earlier lots of ascites, suggesting possible modification of antibody during storage. IEF affinity immunoblot analysis is a useful tool for monitoring MoAb pI microheterogeneity as an indicator of antibody quality without the need for isolation of monoclonal antibody from culture medium or ascites.

Animals↗
PubMed1987

Analysis of immunoglobulin G antibody responses after administration of live and inactivated influenza A vaccine indicates that nasal wash immunoglobulin G is a transudate from serum.

Following intranasal administration of live influenza A virus vaccine or parenteral inoculation of inactivated influenza virus vaccine, immunoglobulin antibody to the influenza virus hemagglutinin was detected in nasal wash specimens from adult volunteers. Several observations supported the suggestion that this immunoglobulin G hemagglutinin nasal wash antibody appeared to be mainly derived from the serum by a process of passive transudation.

Adult↗
PubMed1987

Differential immunoglobulin G subclass antibody titers to respiratory syncytial virus F and G glycoproteins in adults.

Two respiratory syncytial virus glycoproteins, F and G, which differ substantially in the amount of glycosylation were used as antigens in an enzyme-linked immunosorbent assay to determine immunoglobulin G (IgG) subclass titers in 30 experimentally infected healthy adults. The titers of antibodies to the F glycoprotein achieved in postinfection sera were highest in the IgG1 subclass, whereas those to the G glycoprotein were highest and comparable in the IgG1 and IgG2 subclasses. The high IgG2 response to the G glycoprotein suggests that it is seen by the immune system as a polysaccharide antigen, a hypothesis consistent with its large carbohydrate content.

Antibodies, Viral↗
PubMed1987

Subclass restriction of human enterovirus antibodies.

We studied antibodies to enteroviruses in four groups of serum specimens: those from healthy adults, cord blood specimens, serum specimens known to contain immunoglobulin M (IgM) to coxsackie B (CB) viruses by radioimmunoassay, and serum specimens from children with symptomatic enteroviral infections. Enzyme-linked immunosorbent assays (ELISAs) were developed to detect the IgG class- and subclass (IgG1, IgG2, IgG3, and IgG4)-specific responses to CB3. The CB3 virus ELISA was not type specific. There was very poor correlation between CB3 virus neutralizing titer and IgG anti-CB3 virus ELISA results, indicating that antibodies to heterologous picornaviruses cross-react with CB3 virus in the assay. All serum specimens tested except one were IgG positive for CB3 virus. All 32 cord serum specimens were positive for IgG1 and IgG3. No enterovirus-specific IgG2 or IgG4 was detected in any serum specimen tested. Most serum specimens from the IgM-positive group, healthy adults, and children with enterovirus infections were positive for IgG1 and IgG3. Class and subclass antibody titers remained constant over time. IgG antibodies to enteroviruses appear to be restricted to the IgG1 and IgG3 subclasses. This pattern is similar to results obtained by other investigators evaluating IgG subclass antibodies to protein antigens.

Adult↗
PubMed1986

Use of monoclonal antibodies to quantify subclasses of human IgG. II. Enzyme immunoassay to define antigen specific (anti-filarial) IgG subclass antibodies.

Because of their single epitope specificity, monoclonal antibodies (Mcabs) may perform with different levels of efficiency in immunoassays depending on the accessibility of the particular epitope recognized. In order to develop assays capable of detecting specific antibodies of each of the four human IgG subclasses, we have evaluated by ELISA the performance characteristics of a panel of Mcabs raised to the subclass proteins. At least one Mcab to each of the four subclasses was identified that was specific in its ability to capture its own relevant IgG subclass without any associated light chain, allotype or isoallotype activity and that was able to function effectively as a probe in an optimized, quantitative ELISA. When IgG subclass antibodies were measured in sera from patients with filariasis using specific filarial antigen, the sensitivities of each subclass antibody assay varied; for IgG1 and IgG4 antibodies the sensitivity of detection was 50 ng/ml and for IgG2 and IgG3, 10 ng/ml. The potency of the Mcab, determined by its titration for use as a probe, did not correlate with the sensitivity of the assay. These Mcabs were also capable of defining IgG subclass antibody responses qualitatively in immunoblot analyses with little or no non-specific binding. The availability of such highly characterized Mcabs for use in quantitative and qualitative definition of specific IgG subclass antibody responses should greatly improve our detection and subsequent understanding of the role of these IgG subclasses in various disease states.

Antibodies↗
PubMed1986

Use of monoclonal antibodies to quantify subclasses of human IgG. I. Development of two-site immunoenzymometric assays for total IgG subclass determinations.

Dissection of the IgG antibody response into its subclass components has been difficult largely because of the lack of adequate supplies of specific reagents. The development of monoclonal antibodies (Mcab) promises to overcome this problem, but the use of such antibodies has certain inherent problems. It has been shown recently that Mcabs which were avid, potent and specific for well defined epitopes may partially or completely lose their activity depending on the assay system in which they were used. In order to identify Mcabs that would be specific and useful as capture antibodies in a simple two-site enzymometric assay, a panel of 18 Mcabs was screened and one Mcab to each of the four IgG subclasses was identified for quantitation of subclass levels in human serum.

Antibodies, Monoclonal↗
PubMed1986

Serum immunoglobulin G antibody subclass responses to respiratory syncytial virus F and G glycoproteins after primary infection.

Because the immunoglobulin G (IgG) response to carbohydrate antigens is typically from the IgG2 subclass and the IgG response to protein antigens is typically from the IgG1 and sometimes the IgG3 subclass, two respiratory syncytial virus glycoproteins, F and G, which differ substantially in the amount of glycosylation, were used as antigens in an enzyme-linked immunosorbent assay to determine IgG subclass responses in 20 infants and young children with naturally acquired respiratory syncytial virus infection. Both glycoproteins elicited primarily IgG1 and IgG3 responses, indicating that the protein moieties of the glycoproteins may be immunodominant in this age group.

Antigens, Viral↗
PubMed1986

Isotype-specific enzyme immunoassay for influenza antibody with monoclonal antibodies to human immunoglobulins.

Mouse monoclonal antibodies specific for human immunoglobulin isotypes were investigated for use in an isotype-specific enzyme immunoassay for detection of antibody to influenza type A hemagglutinin (H1 and H3). The monoclonal antibody reagents were compared with isotype-specific, hyperimmune rabbit antisera from the National Institutes of Health. Endpoint titers for immunoglobulin G (IgG) obtained with the two reagents were within fourfold of each other 84% of the time (79 of 84) and within eightfold of each other 95% of the time (89 of 94). Regression analysis of the data gave a multiple correlation coefficient (r2) of 0.77 and a Spearman rank value of 0.83 (P less than 0.001). For IgA reagents, endpoint titers agreed within fourfold 77% of the time (88 of 114) and within eightfold 92% of the time (105 of 114). The r2 was 0.73, and Spearman rank was 0.83 (P less than 0.001). IgM antibody was detected in only 17 of 114 sera by either monoclonal or polyclonal reagents. Of these sera, 14 (82%) gave titers with the two reagents that were within fourfold of each other. A similar number of fourfold titer rises were detected with each reagent in paired sera showing hemagglutination inhibition titer rises. Monoclonal antibody reagents detected 27 IgA, 29 IgG, and 6 IgM rises, while polyclonal antisera detected 26 IgA, 31 IgG, and 7 IgM rises. These results show that monoclonal antibodies specific for human immunoglobulin isotypes are suitable as reagents for diagnostic assays. The advantages of monoclonal antibodies are their high degree of specificity and the ability to be standardized and produced in unlimited quantities. Moreover, the availability of immunoglobulin subclass- and allotype-specific monoclonal antibodies will enable a more detailed analysis of the antibody response to influenza as well as other infectious agents.

Animals↗
PubMed1985

Evaluation of monoclonal antibodies having specificity for human IgG sub-classes: results of an IUIS/WHO collaborative study.

Seventy-four monoclonal antibodies (McAb) of putative specificity for human IgG (11), the IgG sub-classes (59) or Gm allotypes (4) have been evaluated for reactivity and specificity in eight laboratories employing different assay techniques or protocols. For the IgG, IgG3, IgG4, G1m(f) and G3m(u) specificities McAb have been produced that can be satisfactorily applied in most methodologies employed and have potential as reference reagents. The IgG1 and particularly IgG2 specificities proved problematical with all McAb evaluated demonstrating apparent assay restriction and whilst performing well in some assays proved to be poor or inactive reagents in others. However, the study identifies McAb individually suited to application within most commonly employed methodologies. Epitope display is the probable variability rather than capricious behaviour by the McAb. IgG1 and IgG2 were the least immunogenic of the sub-class proteins and there is evidence that epitope display is influenced by the physical and chemical procedures used to immobilize or fix antigen - a common requirement in the assay systems studied.

Antibodies, Monoclonal↗
PubMed1985

IgG subclass antibodies to herpes simplex virus.

Several mouse monoclonal antibodies specific for human IgG1, IgG2, IgG3, and IgG4 were evaluated by enzyme-linked immunosorbent assay to detect human IgG subclass antibodies to herpes simplex virus (HSV) antigens. The variable results with different monoclonal antibodies point to the need for well-characterized reagents in the study of antibody responses to infectious agents. The 204 sera tested were obtained from 157 patients with various forms of clinically manifest HSV infections and from several controls. IgG1 antibodies were demonstrated in almost all HSV-infected subjects and were the first antibodies to appear in primary genital infections. IgG2, IgG3, and IgG4 antibodies were detected in acute-phase sera, most often in patients with recurrent genital herpes but in none of those with primary infections. IgG4 antibodies occurred significantly more frequently in sera from men than in those from women with recurrent genital infections.

Adult↗
PubMed1985

Monoclonal antibody-based solid-phase immunoenzymometric assays for quantifying human immunoglobulin G and its subclasses in serum.

We developed quantitative immunoenzymometric assays for human IgG and its subclasses by using monoclonal antibodies, an avidin-biotin detection system and, as the calibrant, the U.S. National Reference Preparation for Specific Human Proteins. The assays are sensitive (detecting as little as 6 micrograms/L), precise (average inter-assay CV less than 11%), and vary linearly with concentrations over a five- to 10-fold range, depending on the monoclonal antibody. We evaluated 22 different monoclonal antibodies, many of which remained highly reactive when immobilized in wells of microtiter plates coated with bovine serum albumin-glutaraldehyde to "capture" total IgG or subclasses of IgG in the sample. We demonstrated the specificity of the most reactive antibodies by using a panel of 20 purified myeloma proteins. The sum of IgG subclass concentrations correlated well (r = 0.84, p less than 0.001) with the total IgG measured in sera from 63 apparently healthy adults (26 men, 37 women). We estimated 95 percentile reference intervals for the immunoglobulins in these subjects and determined the following mean percentage distributions of IgG subclasses: IgG1 49, IgG2 33, IgG3 9, and IgG4 7. The availability of these assays should facilitate studies of the clinical significance of the subclasses.

Adult↗
PubMed1985

Immunoassay for IgG rheumatoid factor with a murine monoclonal anti-Fd antibody.

Conventional radioimmunoassays for IgG rheumatoid factors (RF) detect the binding of IgG RF (or their F(ab')2 fragments) to solid-phase human IgG Fc fragments or rabbit IgG. Binding is detected with a radiolabeled antibody that is IgG-specific but is nonreactive with human Fc (i.e., an alpha-Fd reagent) or with rabbit IgG. Anti-Fd reagents are quite laborious to produce. We developed a murine monoclonal alpha-Fd antibody, confirmed its specificity, and compared it with conventional rabbit alpha-Fd in the IgG RF radioimmunoassay. We also adapted the assay to an enzyme-linked assay with both human Fc and rabbit IgG as antigen. We compared the four assay methods with each other and examined their relationship to certain clinical and laboratory features of rheumatoid arthritis.

Animals↗