DNA microarray analysis of the uninoculated eye following anterior chamber inoculation of HSV-1.
OBJECTIVE
To use DNA microarray to analyze the expression patterns of genes in the uninoculated eye following uniocular anterior chamber inoculation of HSV-1.


METHODS
On Day 9 following inoculation of 2 x 10( 4) PFU of HSV-1 (KOS strain) or an equivalent volume of tissue culture medium into one anterior chamber of BALB/c mice, the uninoculated eyes were enucleated, pooled, and total RNA was isolated. cDNA was synthesized from the total RNA. The gene expression patterns were inferred based on the hybridization intensities of the probes on the cDNA array. The hybridization signals were globally normalized and filtered. The data were analyzed using hierarchical and gene tree clustering algorithms. Additional uninoculated eyes collected on Day 9 p.i. were stained for F4/80 and CD19.


RESULTS
Compared with the uninoculated eye of control mice, 3800 genes were upregulated at least twofold in the contralateral eye of HSV-1-infected mice. Among the 10 most upregulated genes, T cell-specific protein, MHC II antigen A, and MHC II k region locus 2 were upregulated 179-, 164-, and 162-fold, respectively. Ten T-cell receptor-related genes, 61 cytokine and chemokine genes, and 16 MHC genes were upregulated. Furthermore, 11 immunoglobulin and B cell genes and 11 macrophage-related genes were also upregulated. F4/80+ and CD19+ cells were observed on Day 9 p.i.


CONCLUSIONS
The DNA microarray results support the idea that T cells and immunomodulatory factors (cytokines, chemokines) are likely to be involved in HSV-1 retinitis. These results also suggest that B cells and/or macrophages play a role in the pathogenesis of HSV-1 retinitis.
Zheng
Mei
M
Department of Cellular Biology, Medical College of Georgia, Augusta, GA 30912, USA.
Qian
Hailun
H
Joshi
Ratan Mani
RM
Nechtman
John
J
Atherton
Sally S
SS
D000818:Animals; D000867:Anterior Chamber / Q000821:virology*; D018941:Antigens, CD19 / Q000378:metabolism; D000943:Antigens, Differentiation / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology; D015828:Eye Infections, Viral / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D006561:Herpes Simplex / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D018259:Herpesvirus 1, Human / Q000502:physiology*; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000302:isolation & purification; D015882:Retinal Necrosis Syndrome, Acute / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D013601:T-Lymphocytes / Q000276:immunology; D015854:Up-Regulation
D018941:Antigens, CD19; D000943:Antigens, Differentiation; C036763:monocyte-macrophage differentiation antigen; D012313:RNA