Inhibition of collagen-induced arthritis by DNA vaccines encoding TCR Vbeta5.2 and TCR Vbeta8.2.

Abstract

BACKGROUND:

Arthritogenic T lymphocytes with common T cell receptor (TCR) Vbeta clonotypes, infiltrating in the articulars of rheumatoid arthritis (RA) patients, play a central role in the pathogenesis of RA. TCR Vbeta5.2 and TCR Vbeta8.2 are the main pathogenic T cell clonotypes in the course of collagen-induced arthritis (CIA) progression in Lewis rats. To investigate a TCR-based immunotherapy for RA, we constructed recombinant DNA vaccines encoding TCR Vbeta5.2 and TCR Vbeta8.2, and evaluated the inhibitive effects of the two vaccines on CIA rats.

METHODS:

Genes encoding TCR Vbeta5.2 and TCR Vbeta8.2 were amplified by RT-PCR from spleen lymphocytes of Lewis rats and cloned into the eukaryotic expression vector pTargeT. The expression of vaccines was confirmed by RT-PCR and immunohistochemistry. The inhibitive effects of the vaccines on articulars of CIA rats were assessed with arthritis index evaluation and histology. Interferon gamma (IFN-gamma) and interleukin (IL)-4 production by spleen lymphocytes were tested with enzyme-linked immunospot assay (ELISPOT) technique, the changes in peripheral CD4(+) and CD8(+) lymphocyte populations were tested by flow cytometry, and the level of anti-CII antibody in serum was assayed by enzyme-linked immunosorbent assay (ELISA).

RESULTS:

Recombinant DNA vaccines pTargeT-TCR Vbeta5.2 and pTargeT-pTCR Vbeta8.2 were successfully constructed. Both vaccines inhibited CIA, which alleviated the arthritis index score (P < 0.05), decreased the level of IFN-gamma (P < 0.05), and reduced the ratio of CD4(+)/CD8(+) lymphocytes (P < 0.05) and the anti-CII antibody in serum (P < 0.05). In addition, the histological change in DNA-vaccinated rats was less serious than CIA rats. Compared to pTCR Vbeta 8.2 and pTCR Vbeta 5.2 groups, the group that was injected with a combination of the two vaccines showed stronger inhibitive effects on CIA than either individual vaccine.

CONCLUSION:

The recombinant plasmids pTargeT-TCR Vbeta5.2 and pTargeT-TCR Vbeta8.2 have obvious inhibatory effects on CIA rats and better effects could be achieved when the vaccines were used in combination.

• Authors: Ping-ling Ge, Li-ping Ma, Wei Wang, Yun Li, Wen-ming Zhao
• Journal: Chinese medical journal (Chin Med J (Engl)) Vol. 122 Issue 9 Pg. 1039-48 (May 05 2009) ISSN: 2542-5641 [Electronic] China
• PMID: 19493438 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Peptide Fragments
• Receptors, Antigen, T-Cell, alpha-beta
• T cell receptor peptide Vbeta5.2
• T-cell receptor Vbeta 8.2
• Vaccines, DNA
• Interleukin-4
• Interferon-gamma

Topics

• Animals
• Arthritis, Experimental (metabolism, prevention & control)
• CD4-Positive T-Lymphocytes (drug effects)
• CD8-Positive T-Lymphocytes (drug effects)
• Enzyme-Linked Immunosorbent Assay
• Female
• Flow Cytometry
• Immunohistochemistry
• Interferon-gamma (metabolism)
• Interleukin-4 (metabolism)
• Muscles (drug effects, metabolism)
• Peptide Fragments (antagonists & inhibitors)
• Rats
• Rats, Inbred Lew
• Receptors, Antigen, T-Cell, alpha-beta (antagonists & inhibitors)
• Reverse Transcriptase Polymerase Chain Reaction
• Vaccines, DNA (pharmacology)