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Identification of a CD4 variant in Microminipigs not detectable with available anti-CD4 monoclonal antibodies
Tatsuya Matsubara1, Naohito Nishii2, Satoshi Takashima3
1United Graduate School of Veterinary Sciences, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan.
Abstract:
The Microminipig is an extra-small sized novel miniature pig developed in Japan. In the process of peripheral blood mononuclear cells analysis by flow cytometry, CD4+ cells could not be detected in some pigs with an anti-pig CD4 antibody (clone 74-12-4), or in some pigs with two other antibodies from different clones (MIL17 and PT90A). In a herd of 178 Microminipigs, 87 pigs (48.9%) were reactive with the anti-CD4 antibody (designated as CD4.A), and 91 pigs (51.1%) were non-reactive (designated as CD4.B). The CD4 types of piglets delivered from parents with CD4.A were CD4.A or CD4.B, and piglets delivered from parents with CD4.B were only CD4.B. This implies that the CD4.A pigs were homozygous for CD4.A or heterozygous for CD4.A and CD4.B, and the CD4.B pigs were homozygous for CD4.B. The CD4.B trait might be recessive. Significant differences could not be found in the percentage of CD3+ and CD8+ cells in whole lymphocytes between CD4.A and CD4.B animals. In the profile of CD4.B pigs, CD4+CD8+ T cells appeared to be detected in the CD4-CD8+ T cell region because the CD8 dull T cell population was observed. Thus, we considered that the CD4 molecules may be expressed on helper T cells, but the CD4 expressing cells could not be detected with the three anti-pig CD4 antibodies. Clinical abnormalities have not been observed in CD4.B pigs. Significant differences were not observed in immunoglobulin concentrations between CD4.A and CD4.B, though lower tendency was observed in plasma IgM concentrations from CD4.B pigs >36-months-old. These results imply that the CD4.B does not affect basic humoral immunity in vivo.
Insights
A novel Microminipig trait, CD4.B, prevents CD4+ T cell detection via flow cytometry in over half the herd. This trait appears recessive and does not impact basic humoral immunity or cause clinical abnormalities.
Area of Science:
- Veterinary Immunology
- Animal Genetics
- Flow Cytometry Applications
Background:
- The Microminipig, a novel miniature pig model developed in Japan, is used for biomedical research.
- Flow cytometry analysis of peripheral blood mononuclear cells revealed an unexpected inability to detect CD4+ cells in some Microminipigs using specific anti-pig CD4 antibodies.
Purpose of the Study:
- To investigate the genetic basis and immunological implications of a CD4+ T cell detection anomaly in Microminipigs.
- To characterize the CD4.A and CD4.B phenotypes and their potential impact on immune function.
Main Methods:
- Analysis of peripheral blood mononuclear cells from 178 Microminipigs using flow cytometry with multiple anti-pig CD4 antibodies.
- Pedigree analysis to determine the inheritance pattern of the CD4.A and CD4.B traits.
- Comparison of CD3+, CD8+, and immunoglobulin levels between CD4.A and CD4.B pigs.
Main Results:
- A CD4.B phenotype, characterized by the non-reactivity of CD4+ T cells to standard antibodies, was identified in 51.1% of the Microminipig herd.
- The CD4.B trait demonstrated a recessive inheritance pattern, suggesting homozygous recessive individuals.
- No significant differences in CD3+ or CD8+ T cell percentages were observed between CD4.A and CD4.B pigs, and basic humoral immunity, including immunoglobulin levels, was largely unaffected, except for a tendency towards lower IgM in older CD4.B pigs.
Conclusions:
- The CD4.B trait in Microminipigs represents a novel genetic variation affecting CD4 molecule detection rather than absence.
- This variation does not appear to compromise fundamental immune responses or cause clinical issues in the studied population.
- Further research may explore the precise molecular mechanism of CD4 expression or detection in CD4.B Microminipigs.
