Document Type

Article

Journal/Book Title/Conference

Indian Journal of Experimental Biology

Volume

56

Publisher

NISCAIR-CSIR, India

Publication Date

8-2018

First Page

547

Last Page

555

Creative Commons License

Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Abstract

The non-classical MHC class I (MHC-Ib) proteins are important modulators of immune system during pregnancy favoring survival of the fetus. Contrary to ubiquitously expressed classical MHC-I proteins, MHC-Ib proteins are oligomorphic, and expressed in specific cell/tissue types thus minimizing maternal immune-mediated rejection of fetal-allograft and a successful pregnancy. A unique characteristic of MHC-Ib glycoproteins is expression of surface and soluble isoforms due to alternative splicing phenomenon. Bovine fetal trophoblast cells, during the third trimester of pregnancy, express non-classical bovine leukocyte antigen class Ib (BoLA-Ib) antigens. BoLA-NC3*00101, is a non-classical class I allele from cattle AH11 haplotype and is expressed as cell surface isoform. However, lack of monoclonal antibody (mAb) hinders the development of specific assay to detect the soluble/secreted BoLA-I antigens released by fetal trophoblast cells. The objective of this study is to synthesize mAb specific to NC3*00101 molecule to develop an isotype-matched ELISA to assess BoLA-I protein(s). We demonstrated that majority of NC3*00101 hybridoma-supernatants were low-titer and showed inappreciable reactivity in ELISA and flow cytometry assays. We identified a clone of hybridoma (NC3-3*2) that secreted mAb specific to BoLA-NC3*00101 as demonstrated with flow cytometry. NC3-3*2 clone secreted supernatant that captured BoLA-NC3*00101 protein in ELISA. Unfortunately, despite several efforts we could not recover the cryopreserved hybridoma. Non-recoverability and instability, thus, limited production of NC3*00101-mAbs. This study provides the evidence that mice immunized with bovine class Ib proteins elicit a specific immunogenic response and warrants future studies into generation of stable hybridoma secreting antibodies against BoLA-Ib proteins using robust methods of fusion and cryopreservation of hybridomas.

Included in

Dairy Science Commons

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.