Repertoire Breadth of Human CD4+ T Cells Specific for HIV gp120 and p66 (Primary Antigens) or for PPD and Tetanus Toxoid (Secondary Antigens)

Pira, Giuseppina Li; Oppezzi, L.; Seri, Marco; Westby, Michael; Caroli, Francesco; Fenoglio, Daniela; Lancia, F.; Ferraris, Anna Maria; Bottone, Laura; Valle, Maurizio; Kunkl, Annalisa; Romeo, Giovanni; Dalgleish, Angus; Manca, Fabrizio

doi:10.1016/s0198-8859(98)00004-4

Cited by 18 publications

(17 citation statements)

References 36 publications

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“…However, because of the large number of epitopes present within TT, a heterogeneous response such as that observed here is reasonably expectable (16,36). In addition, previous studies strongly support our observation, i.e., the demonstration of a broad clonal heterogeneous response to a recall response to TT, with a high frequency of specific precursors, as opposed to a rather oligoclonal T-cell response to primary antigens (HIV gp120 or HIV p66) (31). Taken together, as supported by cloning and TCR spectratyping, our phenotypic approach demonstrated that in this model of TT immunization a substantial proportion of antigen-specific CD4 ϩ T cells were included within the CCR7 ϩ/Ϫ CD69 ϩ IL-2 ϩ CD4 ϩ T-cell subpopulations (up to 50%).…”

Section: Discussionsupporting

confidence: 87%

Ex Vivo Monitoring of Antigen-Specific CD4⁺T Cells after Recall Immunization with Tetanus Toxoid

Barbey

Pradervand

Barbier

et al. 2007

Clin Vaccine Immunol

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To monitor antigen-specific CD4؉ T cells during a recall immune response to tetanus toxoid (TT), a sequential analysis including ex vivo phenotyping and cytokine flow cytometry, followed by cloning and T-cell-receptor (TCR) spectratyping of cytokine-positive CD4 ؉ T cells, was performed. Grossly, twice as many TT-specific CD4 ؉ T-cell clones, ex vivo derived from the CCR7 ؉ subsets appears to target the bulk of antigen-specific T cells and to reach an analytical power sufficient to adequately delineate in field trials the profile of the antigen-specific response to vaccine.Phenotypic characterization of antigen-specific CD4 ϩ T cells with distinct functional properties represents currently a major challenge in human applied experimental immunology. Ex vivo analysis of these cell populations is critical, since in vitro culture may alter composition and functional properties of the populations of interest. If major histocompatibility complex (MHC) class I tetramer/peptide complexes did greatly facilitate the analysis of antigen specific CD8 ϩ T cells, the use of MHC class II tetramer/peptide has been hampered by the complexity of MHC class II tetramer production, by the usually low CD4 ϩ T-cell precursor frequency, by an often-degenerated peptide epitope specificity, and by non-MHC class II moleculerestricted allelic expression (12,15,24,35,36). As an alternative approach to circumvent these difficulties, we here applied cytokine flow cytometry (9, 11), now extensively used to define ex vivo frequencies and phenotypic and functional characteristics of viral antigen-specific CD4 ϩ T cells (8,22,43), coupled to T-cell-receptor (TCR) spectratype analysis. Indeed, characterization of the unique complementary determining region 3 (CDR3) sequence expression of CD4 ϩ (or CD8 ϩ ) T-cell clones appears to be a sensitive method for monitoring qualitative and quantitative T-cell responses during natural infection or vaccination (21,23,45,48).In the present study, tetanus toxoid (TT) was used as a model vaccine antigen that likely shares common functional characteristics with other nonviral antigens, such as a low frequency of specific CD4 ϩ T cells in the periphery at steady state and a specific CD4 ϩ T-cell response to multiple epitope peptides presented by MHC class II molecules (16,36,40). Using ex vivo cell phenotyping and cloning in combination with TCR spectratype analysis of cytokine-positive CD4 ϩ T cells, we sought to establish experimental conditions to track ex vivo antigenspecific CD4 ϩ T cells in conditions that are likely to reflect clinical situations such as vaccination, nonviral pathogen infections, or allergic diseases.

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Section: Discussionsupporting

confidence: 87%

Ex Vivo Monitoring of Antigen-Specific CD4⁺T Cells after Recall Immunization with Tetanus Toxoid

Barbey

Pradervand

Barbier

et al. 2007

Clin Vaccine Immunol

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“…Two donors, well characterized for their cellular immune responses to the test antigens and from whom HIV-specific T-cell lines could be generated in spite of their HIV seronegative status (Manca et al, 1991;Li Pira et al, 1998), volunteered for these experiments. Venous heparinized blood was diluted 1/2 with PBS and 20 ml were layered over 10 ml Lymphocyte Separation Medium (LSM, BioWhittaker) in 50 ml tubes.…”

Section: Pbmc Preparationmentioning

confidence: 99%

“…HIV specific CD4 T-cell lines were obtained from two healthy, seronegative donors, as previously described in detail (Manca et al, 1991(Manca et al, , 1995Li Pira et al, 1998). One line was specific for HIV envelope glycoprotein gp120 and recognized the immunodominant peptide 191-205 (PAGFAILKCNNTFNY).…”

Section: Generation Maintenance and Use Of Hiv Specific Cd4 T-cell Lmentioning

confidence: 99%

Lymphocyte proliferation specific for recall, CMV and HIV antigens in miniaturized and automated format

Pira

Starc

Conforti

et al. 2012

Journal of Immunological Methods

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“…Even though the total number of residual specific cells may be similar in the two instances, in case of immunoreconstitution (ie recovery of CD4 counts) horizontal depletion is compatible with restoration of a clonally heterogeneous repertoire, whereas vertical depletion is compatible with a skewed repertoire recovery only. Skewing may occurr as a consequence of in vitro culture 96,97 or when therapeutic genes are artificially introduced. 70 Therefore, monitoring of clonal heterogeneity of antigenspecific T cell lines should be performed according to TCR BV gene family usage and spectrotyping.…”

Section: Clonal Heterogeneity Of Antigen-specific Cd4 T Cellsmentioning

confidence: 99%

“…This method can identify in vitro fewer than 40 clonotype positive cells out of one million negative cells. 96 TCR BV gene usage with sequencing of the PCR products was reported as a way to monitor CD8 clones specific for CMV. 99 A similar approach can be adopted to trace antigen-specific CD4 cells independently of the marker gene utilized.…”

Section: Clonotypic Tracingmentioning

confidence: 99%

Genetically modified immunocompetent cells in HIV infection

Palù

Pira²,

Gennari

et al. 2001

Gene Ther

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Even in the era of highly active antiretroviral therapy (HAART), gene therapy (GT) can remain a promising approach for suppressing HIV infection, especially if complemented with other forms of pharmacological and immunological intervention. A large number of vectors and targets have been studied. Here we discuss the potential of genetically treated, antigen-specific immunocompetent cells for adoptive autologous immunotherapy of HIV infection. Cellu-

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Repertoire Breadth of Human CD4+ T Cells Specific for HIV gp120 and p66 (Primary Antigens) or for PPD and Tetanus Toxoid (Secondary Antigens)

Cited by 18 publications

References 36 publications

Ex Vivo Monitoring of Antigen-Specific CD4⁺T Cells after Recall Immunization with Tetanus Toxoid

Ex Vivo Monitoring of Antigen-Specific CD4⁺T Cells after Recall Immunization with Tetanus Toxoid

Lymphocyte proliferation specific for recall, CMV and HIV antigens in miniaturized and automated format

Genetically modified immunocompetent cells in HIV infection

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Repertoire Breadth of Human CD4+ T Cells Specific for HIV gp120 and p66 (Primary Antigens) or for PPD and Tetanus Toxoid (Secondary Antigens)

Cited by 18 publications

References 36 publications

Ex Vivo Monitoring of Antigen-Specific CD4+T Cells after Recall Immunization with Tetanus Toxoid

Ex Vivo Monitoring of Antigen-Specific CD4+T Cells after Recall Immunization with Tetanus Toxoid

Lymphocyte proliferation specific for recall, CMV and HIV antigens in miniaturized and automated format

Genetically modified immunocompetent cells in HIV infection

Contact Info

Product

Resources

About

Ex Vivo Monitoring of Antigen-Specific CD4⁺T Cells after Recall Immunization with Tetanus Toxoid

Ex Vivo Monitoring of Antigen-Specific CD4⁺T Cells after Recall Immunization with Tetanus Toxoid