{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2022,4,5]],"date-time":"2022-04-05T19:20:12Z","timestamp":1649186412625},"reference-count":21,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2007,4,1]],"date-time":"2007-04-01T00:00:00Z","timestamp":1175385600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Clin Vaccine Immunol"],"published-print":{"date-parts":[[2007,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Cell-mediated immunity, particularly that induced by T cells, is thought to have a key role in controlling infection. The enzyme-linked immunospot (ELISPOT) assay has been successfully adapted to detect T-cell immune response to a variety of pathogens. However, it still remains a challenge to detect antigen-specific T cells when the numbers of circulating cells are low, such as in a local cervical infection caused by genital human papillomavirus (HPV). The goal of this study was to develop a protocol for enhanced detection of HPV-specific CD8\n            <jats:sup>+<\/jats:sup>\n            T cells by examining a number of the variables involved in performing an ELISPOT assay. Since blood samples consistently positive for HPV-specific T cells are difficult to obtain, previously described human papillomavirus type 16 (HPV16) E6 52-61 (FAFRDLCIVY)-specific T-cell clone cells (13) seeded in peripheral blood mononuclear cells from an HLA-B57-positive blood donor were used. The variables examined were the amounts of primary and secondary anti-gamma interferon antibodies, amounts of antigen-presenting monocytes and recombinant vaccinia virus expressing the HPV16 E6 protein, and amounts of exogenous cytokines added (recombinant human interleukin-2 [rhIL-2] and rhIL-7). The amounts of antigen-presenting monocytes, followed by the concentration of exogenous rhIL-2, had the most pronounced and significant effects in enhancing sensitivity of the ELISPOT assay. Blood samples from six patients being monitored for abnormal Pap smear results and from 12 healthy volunteers were examined using the enhanced conditions.\n          <\/jats:p>","DOI":"10.1128\/cvi.00460-06","type":"journal-article","created":{"date-parts":[[2007,2,22]],"date-time":"2007-02-22T01:47:02Z","timestamp":1172108822000},"page":"362-368","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Detection of Human Papillomavirus Type 16-Specific T Lymphocytes by a Recombinant Vaccinia Virus-Based Enzyme-Linked Immunospot Assay"],"prefix":"10.1128","volume":"14","author":[{"given":"Kevin H.","family":"Kim","sequence":"first","affiliation":[{"name":"Departments of Dermatology"},{"name":"Obstetrics and Gynecology"},{"name":"Pathology, University of Arkansas for Medical Sciences, Little Rock, Arkansas"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"William","family":"Greenfield","sequence":"additional","affiliation":[{"name":"Departments of Dermatology"},{"name":"Obstetrics and Gynecology"},{"name":"Pathology, University of Arkansas for Medical Sciences, Little Rock, Arkansas"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ezekiel","family":"Shotts","sequence":"additional","affiliation":[{"name":"Departments of Dermatology"},{"name":"Obstetrics and Gynecology"},{"name":"Pathology, University of Arkansas for Medical Sciences, Little Rock, Arkansas"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mayumi","family":"Nakagawa","sequence":"additional","affiliation":[{"name":"Departments of Dermatology"},{"name":"Obstetrics and Gynecology"},{"name":"Pathology, University of Arkansas for Medical Sciences, Little Rock, Arkansas"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.274.5284.94"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00774-06"},{"key":"e_1_3_2_4_2","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.jim.2005.01.003","volume":"298","year":"2005","unstructured":"Buseyne, F., A. Catteau, D. Scott-Algara, B. Corre, F. Porrot, C. Rouzioux, S. Blanche, and Y. Riviere. 2005. A vaccinia-based elispot assay for detection of CD8+ T cells from HIV-1 infected children. J. Immunol. Methods298:105-118.","journal-title":"J. Immunol. Methods"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.80.10.4717-4728.2006"},{"key":"e_1_3_2_6_2","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/S0022-1759(02)00275-2","volume":"270","year":"2002","unstructured":"Jennes, W., L. Kestens, D. F. Nixon, and B. L. Shacklett. 2002. Enhanced ELISPOT detection of antigen-specific T cell responses from cryopreserved specimens with addition of both IL-7 and IL-15\u2014the Amplispot assay. J. Immunol. Methods270:99-108.","journal-title":"J. Immunol. Methods"},{"key":"e_1_3_2_7_2","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/0022-1759(93)90158-4","volume":"159","year":"1993","unstructured":"Jung, T., U. Schauer, C. Heusser, C. Neumann, and C. Rieger. 1993. Detection of intracellular cytokines by flow cytometry. J. Immunol. Methods159:197-207.","journal-title":"J. Immunol. Methods"},{"key":"e_1_3_2_8_2","doi-asserted-by":"crossref","first-page":"1303","DOI":"10.1172\/JCI12433","volume":"107","year":"2001","unstructured":"Kaul, R., T. Dong, F. A. Plummer, J. Kimani, T. Rostron, P. Kiama, E. Njagi, E. Irungu, B. Farah, J. Oyugi, R. Chakraborty, K. S. MacDonald, J. J. Bwayo, A. McMichael, and S. L. Rowland-Jones. 2001. CD8+ lymphocytes respond to different HIV epitopes in seronegative and infected subjects. J. Clin. Investig.107:1303-1310.","journal-title":"J. Clin. Investig."},{"key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"5675","DOI":"10.4049\/jimmunol.175.9.5675","volume":"175","year":"2005","unstructured":"Keating, S. M., P. Bejon, T. Berthoud, J. M. Vuola, S. Todryk, D. P. Webster, S. J. Dunachie, V. S. Moorthy, S. J. McConkey, S. C. Gilbert, and A. V. Hill. 2005. Durable human memory T cells quantifiable by cultured enzyme-linked immunospot assays are induced by heterologous prime boost immunization and correlate with protection against malaria. J. Immunol.175:5675-5680.","journal-title":"J. Immunol."},{"key":"e_1_3_2_10_2","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/S0022-1759(97)00177-4","volume":"210","year":"1997","unstructured":"Lalvani, A., T. Dong, G. Ogg, A. A. Patham, H. Newell, A. V. Hill, A. J. McMichael, and S. Rowland-Jones. 1997. Optimization of a peptide-based protocol employing IL-7 for in vitro restimulation of human cytotoxic T lymphocyte precursors. J. Immunol. Methods210:65-77.","journal-title":"J. Immunol. Methods"},{"key":"e_1_3_2_11_2","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1097\/00002030-199905070-00005","volume":"13","year":"1999","unstructured":"Larsson, M., X. Jin, B. Ramratnam, G. S. Ogg, J. Engelmayer, M. A. Demoitie, A. J. McMichael, W. I. Cox, R. M. Steinman, D. Nixon, and N. 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