Studies of Monoclonal Antibodies Specific for Major Histocompatibility Complex Products of the Rat

Sixteen monoclonal anti-idiotypic antibodies were prepared against RT1 antigen-specific monoclonal antibodies by means of an inhibition assay. Three (110-3, 112-89, and 133-4) were produced against antibody 3-8C2 (19), three (178-60, 181-14, and 184-38) against antibody 11/23/5, and three (144-3, 144-33, and 146-31) against antibody 1-5A4. One monoclonal anti-idiotypic antibody (21-39) was directed against antibody 1-8D2 and six (CE1-3, CE2-1, CE7-2, CE8-46, CE9-35, and CE10-2) against antibody 3-12A1. The anti-idiotypic antibodies were all highly specific for the respective monoclonal antibody used for the induction of the anti-idiotype and did not cross-react with other monoclonal antibodies.     

Studies of Monoclonal Antibodies Specific for Major Histocompatibility Complex Products of the Rat

Polyclonal anti-idiotypic antibodies against monoclonal antibodies with specificity for RT1-encoded antigens were induced in syngeneic, allogeneic, and xenogeneic hosts. The immune response to the idiotypic determinants on monoclonal antibodies was T-cell-dependent. The anti-idiotypic antibodies, independent of whether they were induced in syngeneic, allogeneic, or xenogeneic (after proper absorption) hosts, showed an exquisite specificity for the monoclonal antibody used for the induction. No cross-reactivity with other monoclonal antibodies could be observed.     

Studies of Monoclonal Antibodies Specific for Major Histocompatibility Complex Products of the Rat

Sixteen monoclonal antibodies against antigens coded for by the RT1 complex of the rat have been produced. Fourteen are specific for the a haplotype: six recognize class II and eight class I antigens. Two are specific for the 1 haplotype, one reacting with class I and the other with class II antigens. By means of these monoclonal antibodies four independent clusters of antigens for class I antigens of the a haplotype and three for class II antigens could be defined. The three antigenic sites of class II antigens reside on the same heterodimer. The monoclonals described here are characterized with regard to Ig class and subclass, pI, and complement-activating capacity.     

抗大鼠树突状细胞单克隆抗体的研制及初步鉴定

树突状细胞是一种重要的抗原提呈细胞,在移植排斥反应中发挥重要作用。为了进一步研究该细胞的功能和寻找抗移植排斥反应的更有效途径,我们在分离大鼠树突状细胞获得成功的基础上,应用杂交瘤技术制备出5株抗树突状细胞单克隆抗体,分别命名为WZD1～5。杂交瘤细胞培养上清或腹水对新鲜分离的大鼠树突状细胞均显示不同程度的细胞毒性作用,且与腹腔Mφ及淋巴细胞无明显交叉反应。经间接免疫荧光染色测定,腹水中单抗的效价为2×10~(-2)。     

Persistence and Selectivity of the Immune Phagocytosis Inhibition by Major Histocompatibility Complex Antibodies

Major histocompatibility complex (MHC) antibodies induce immune phagocytosis inhibition (IPI) which lasts for at least 7 days. IPI-inducing antibodies do not inhibit the phagocytosis mediated by the beta-glucan receptor. This corresponds well to recent findings that these antibodies do not interfere with the phagocytosis of deactivated saccharomyces, mediated by the mannose-fucose receptor, or polyacrylic acid particles, also mediated by non-Fc receptors. Substances that interact with certain MHC antigens or with Fc receptors, certain toxins that inhibit surface molecule mobility, and ciclosporin do not cause IPI and do not suppress the induction of IPI by MHC antibodies. These substances are: opioid peptides, insulin, penicillin G, immune complexes, aggregated IgG, Fc fragments, ciclosporin, botulinum C2 toxin, sodium azide. Some lectins and EDTA are inhibitory in a non-selective fashion, since the Fc receptor independent phagocytosis is also abrogated.     

Peptide Dependence of Major Histocompatibility Complex Class II Specific Alloreactive Responses

Splenic cells from transgenic mice, in which a single peptide is complexed to all major histocompatibility complex (MHC) class II molecules, are found to be incapable of triggering primary allogeneic mixed lymphocyte/leucocyte reactions (MLR) when co-cultured with lymphocytes from MHC class II congenic mouse strains. In addition, a single HLA-DR-blocking peptide can completely abrogate the capacity of splenocytes from chimeric HLA-DR/H2-E transgenic mice to stimulate primary MLR of T cells from wild-type mice. These results indicate that the primary alloreactive response is directed against a multitude of peptides presented by allogeneic MHC molecules.     

Monoclonal Antibodies Specific for Sendai Virus

The monoclonal antibodies A and B specific for the HN molecule of Sendai virus were used to induce polyclonal and monoclonal anti-idiotypic antibodies. No response was observed in allogeneic Lewis rats, low responses in syngeneic LOU rats, and high responses in allogeneic BN rats and xenogeneic Balb/c mice. The monoclonals A and B share a similar or identical idiotype, since polyclonal anti-idiotypic antisera to antibody A cross-reacted completely with antibody B and vice versa. The same was found with the three monoclonal anti-idiotypes 1, 2, and 3 elicited in a BN rat or in Balb/c mice. None of the polyclonal or monoclonal anti-idiotypes reacted with other monoclonal antibodies specific for Sendai virus, even when these recognized the same epitope as antibodies A and B. The monoclonal anti-idiotypic antibodies could be used to induce anti-Sendai antibodies in BN rats.     

抗大鼠视网膜神经节细胞诱向因子单克隆抗体的产生及其鉴定

视网膜神经节细胞诱向因子(RGNTF,Retinal ganglion neuronotrop-bic factor)是新发现的一神经生长因子,它能特异性地促进体外培养的视网膜神经节细胞的生长活性。作者用Phast System电泳实验证实,RGNTF的分子量为30kD蛋白质。并用之免疫动物,制备出3株抗RGNTF单克隆抗体,分别命名为A1、D3、E8。经鉴定杂交瘤细胞系染色体大多为端着丝点染色体,并有1～2条中部着丝点染色体。抗体的亚型鉴定A1、D3是IgG3型、E8是IgM型。用ELISA方法测定RGNTF-McAb与相应抗原结合的效价为10~(-4)～10~(-5)结果发现随抗体浓度的增强,抗体与抗原结合率也增加,以E8株结合率最高。本单抗与其它生长因子MGF、EGF无交叉反应,表明其相应抗原是一种新的生长因子。     

Birth of the Major Histocompatibility Complex

Klein J, Sato A. Birth of the Major Histocompatibility Complex. Scand J Immunol 1998;47:199–209     

Monoclonal Antibodies Specific for Sendai Virus I. Production and Characterization of Monoclonal Antibodies

Twelve monoclonal antibodies specific for Sendai virus were prepared by fusing immune LOU rat spleen cells with Y3 or FO myeloma cells. Six antibodies bound the viral glycoprotein HN, and six the viral protein F. Among the six HN-specific monoclonal antibodies, five reacted with the very same epitope and inhibited viral haemagglutination. Two antibodies against the F protein recognized the same epitope, but all the others reacted with different epitopes. All monoclonals were characterized with regard to specificity, biological function, epitope recognition, isotypes, and pI.     

Monoclonal Rat Antibodies to Rat Carcinoembryonic Antigen

An IgM-class rat monoclonal antibody, designated 5B1, was produced against rat carcinoembryonic antigen. This antibody, which can be considered an autoantibody, formed a single precipitation line in a gel diffusion test with an extract of the CEA-positive rat colonic adenocarcinoma, RCA-1. This line gave a reaction of complete identity with a line formed by a rabbit antiserum specific for rat CEA. 5B1 also bound to extracts of the RCA-1 tumor in EIA, but did not bind at all to extracts of the CEA-negative rat colonic adenocarcinoma, RCA-2, nor to extracts of normal rat tissues including liver, kidney and intestine. Inhibition tests showed that the activity of the monoclonal antibody in EIA was abolished by extracts of RCA-1 tumor, but was hardly affected by extracts of the other tissues.     

Monoclonal Antibodies for Specific Detection of Encephalitozoon cuniculi

Seven species-specific monoclonal antibodies (MAbs) were produced against Encephalitozoon cuniculi and characterized. The MAbs were immunoglobulin G, and when used for indirect microimmunofluorescence microscopy and Western immunoblot assays, they detected E. cuniculi originating from clinical samples. They did not cross-react with other Encephalitozoon species (E. intestinalis and E. hellem) or with a collection of gram-negative bacteria, yeast, and other parasites. The MAbs reacted primarily with 121-, 56-, 45-, 43-, and 41-kDa protein epitopes of E. cuniculi. These epitopes were demonstrated to be E. cuniculi species specific by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We developed MAbs to strains of E. cuniculi that can be used successfully to distinguish E. cuniculi from other microsporidial species in cultures established from clinical specimens. These MAbs may provide a specific, simple, rapid, and low-cost tool for the identification and diagnosis of infections due to microsporidia.