ABSTRACT
Recurrent human herpes simplex lesions are infiltrated by macrophages and CD4 and CD8 lymphocytes, which secrete cytokines and chemokines. Vesicle fluid was examined by ELISA for the presence of cytokines and beta (C-C) chemokines. On the first day of the lesion, high concentrations of interleukin (IL)-1beta, and IL-6, moderate concentrations of IL-1alpha and IL-10, and low concentrations of IL-12 and beta chemokines were found; levels of macrophage inflammatory protein (MIP)-1beta were significantly higher than levels of MIP-1alpha and RANTES. At day 3, the concentrations of IL-1beta, IL-6, and MIP-1beta were lower, whereas the levels of IL-10, IL-12, and MIP-1alpha remained similar, and the level of tumor necrosis factor-alpha was now detectable. Herpes simplex virus infection of keratinocytes in vitro stimulated production of beta chemokines followed by IL-12 and then IL-10, IL-1alpha, IL-1beta, and IL-6, indicating a potential role for these events in early recruitment, activation, and interferon-gamma production of CD4 cells in herpetic lesions.
Subject(s)
Chemokines/metabolism , Cytokines/metabolism , Herpes Simplex/immunology , Herpesvirus 1, Human/immunology , Keratinocytes/immunology , Keratinocytes/virology , Adolescent , Adult , Antigens, Viral/immunology , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Cells, Cultured , Chemokine CCL3 , Chemokine CCL4 , Chemokine CCL5/immunology , Chemokine CCL5/metabolism , Culture Media, Conditioned/chemistry , Humans , Interleukin-1/immunology , Interleukin-1/metabolism , Interleukin-10/immunology , Interleukin-10/metabolism , Interleukin-12/immunology , Interleukin-12/metabolism , Interleukin-6/immunology , Interleukin-6/metabolism , Keratinocytes/metabolism , Macrophage Inflammatory Proteins/immunology , Macrophage Inflammatory Proteins/metabolism , Macrophages/immunology , Middle Aged , Pemphigus/virology , Recurrence , Th1 Cells/immunology , Th1 Cells/metabolism , Th2 Cells/immunology , Th2 Cells/metabolism , Tumor Necrosis Factor-alpha/immunology , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Mosaic proteins consist of a group of proteins that may be comprised of one or more types of a variety of different structural modules and have a diverse range of functions. We have examined primary human astrocyte cultures for the presence of three mosaic proteins, B2I and the complement proteins factor H and properdin. Using the polymerase chain reaction and an enhanced chemiluminescence detection technique, we were able to show that mRNA transcripts for each of these proteins are expressed in human astrocytes.
Subject(s)
Astrocytes/metabolism , Complement Factor H/genetics , Glycoproteins/genetics , Nerve Tissue Proteins/genetics , Properdin/genetics , RNA, Messenger/analysis , Base Sequence , Complement Factor H/biosynthesis , DNA/genetics , Gene Expression Regulation , Glycoproteins/biosynthesis , Humans , Luminescent Measurements , Molecular Sequence Data , Nerve Tissue Proteins/biosynthesis , Polymerase Chain Reaction , Properdin/biosynthesis , RNA, Messenger/genetics , Transcription, Genetic , beta 2-Glycoprotein IABSTRACT
Primer sets specific for complement proteins, C3, factor B and factor I, were designed and used to amplify cDNA from cultured human astrocyte mRNA by the polymerase chain reaction. Appropriately sized PCR products of 506 bp, 885 bp and 146 bp, respectively, were generated and specificity was confirmed with Southern blotting using an enhanced chemiluminescence detection system. The sensitivity of detection was high, with amplified product from cDNA of approximately 6250 cells readily visualized. C3 and factor B have previously been reported to be produced by murine astrocytes; however, this is the first report indicating synthesis of C3, factor B and factor I by human astrocytes. These results indicate that PCR is a simple and sensitive technique to detect mRNA transcripts for proteins of the alternative pathway of complement in human astrocytes.
