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Published on: August 13, 2009
Longitudinal analysis of herpes simplex virus-specific CD4+ cell clonotypes in infected tissues and blood
Serge Barcy1, Meei-Li Huang, Lawrence Corey
1Department of Laboratory Medicine, University of Washington, Seattle, Washington 98109-8070, USA. sbarcy@u.washington.edu
Insights
Herpes simplex virus (HSV)-2 infection reveals how T cells persist after genital herpes. Some virus-specific T cells travel to different tissues and blood, while others remain localized to lesions.
Area of Science:
- Immunology
- Virology
- Dermatology
Background:
- Genital herpes simplex virus (HSV)-2 infection provides a model to study immune responses.
- Understanding T cell dynamics during remitting and exacerbating infections is crucial.
Purpose of the Study:
- To investigate the persistence of antigen-specific T cells in genital herpes lesions.
- To analyze the T cell receptor (TCR) repertoire in response to HSV-2 infection.
Main Methods:
- Complementarity-determining region 3 (CDR3) length analysis of TCR beta-chain repertoire.
- TCRBV sequencing and clonal tracking techniques to identify HSV-specific CD4(+) T cells.
- Polymerase chain reaction and liquid hybridization for sensitive clonal detection.
Main Results:
- Herpetic skin lesions showed oligoclonal CDR3 DNA length distributions, indicating T cell expansion.
- Oligoclonal expansions were primarily due to HSV-specific T cell proliferation.
- A novel clonal tracking technique confirmed the persistence of HSV-specific CD4(+) cells.
Conclusions:
- Two distinct patterns of T cell clonal persistence were observed in genital herpes.
- Some long-lasting T cell clones home to multiple epithelia (skin, genital mucosa) and circulate in peripheral blood.
- Other T cell clones detected in lesions were rare or absent in peripheral blood, suggesting localized persistence.
Background:
Genital infection by herpes simplex virus (HSV)-2 offers a unique model for study of the effects of a remitting and exacerbating infection on the survival and persistence of antigen-specific T cells.
Methods:
We used complementarity-determining region 3 (CDR3) length analysis to examine the complete T cell receptor (TCR) beta -chain repertoire in skin-lesion biopsy samples from subjects with genital herpes.
Results:
We found that herpetic skin lesions consistently demonstrated oligoclonal CDR3 DNA length distribution, indicating the presence of T cell expansions. Sequence analysis of representative HSV-specific lesional CD4(+) cell clones and TCR beta -variable (TCRBV) sequencing confirmed that the oligoclonal expansions were largely related to HSV-specific T cell proliferation. To assess the persistence of HSV-specific CD4(+) cells that localize to genital lesions, we developed a sensitive and highly specific clonal tracking technique using a combination of TCRBV-specific polymerase chain reaction, followed by liquid hybridization with clonotype-specific probes.
Conclusion:
Two different patterns of clonal persistence were observed. Some long-lasting clones appear to home to different epithelia, such as skin and genital mucosa, and to circulate in the peripheral blood, whereas others detected in lesions were absent or very rare in the peripheral blood.
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