Identification of viral antigens recognized by ocular infiltrating T cells from patients with varicella zoster virus-induced uveitis.
OBJECTIVE
Varicella zoster virus (VZV) is a common cause of infectious uveitis associated with an intraocular inflammatory response involving virus-specific T cells. In the current study, the functional characteristics and the antigen specificity of VZV-reactive T cells recovered from intraocular fluid (IOF) samples of five patients with VZV were determined.


METHODS
B-cell lines were infected with a comprehensive panel of recombinant vaccinia viruses expressing 11 individual VZV open reading frames (ORFs), or alternatively pulsed with the corresponding peptides to generate antigen-presenting cells (APCs). T-cell responsiveness of the IOF-derived VZV-specific T cells toward APCs was monitored by interferon (IFN)-gamma enzyme-linked immunosorbent spot-forming assays on bulk T-cell cultures and subsequently T-cell clones (TCCs). The cytokine-secretion profile and cytotoxicity of the VZV-specific TCCs was determined by ELISA and flow cytometry, respectively.


RESULTS
T-cell reactivity to VZV proteins encoded by ORF4, -10, -14, -18, -29, -31, -61, -62, -63, -67, and -68 was demonstrated, but specificity varied individually. T-cell epitopes on ORF62 and -68 were delineated. The TCCs secreted IFNgamma, but relatively low levels of interleukin-4 and -5, in response to VZV antigen-expressing APCs. The TCCs induced antigen-specific cytotoxic T-cell activity.


CONCLUSIONS
The results suggest that the intraocular VZV-specific T-cell response in the patients with VZV analyzed is directed to a broad spectrum of VZV antigens, including the latency-associated VZV proteins from ORFs 4, 29, 63, and particularly ORF62. This local T-cell response was in part mediated by cytotoxic CD4(+) T cells with a Th1/0-like effector memory phenotype.
Milikan
Johannes C M
JC
Institute of Virology, Erasmus Medical Center, Rotterdam.
Kinchington
Paul R
PR
Baarsma
G Seerp
GS
Kuijpers
Robert W A M
RW
Osterhaus
Albert D M E
AD
Verjans
Georges M G M
GM
D000293:Adolescent; D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D001082:Aqueous Humor / Q000166:cytology / Q000276:immunology / Q000821:virology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000821:virology; D002461:Cell Line, Transformed; D016207:Cytokines / Q000378:metabolism; D000939:Epitopes; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D005260:Female; D006562:Herpes Zoster / Q000150:complications / Q000276:immunology*; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000821:virology; D018417:Th1 Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism / Q000821:virology*; D014605:Uveitis / Q000276:immunology* / Q000821:virology*
D000956:Antigens, Viral; D016207:Cytokines; D000939:Epitopes; D018984:Epitopes, T-Lymphocyte; D000949:Histocompatibility Antigens Class II