Comparison of overlapping peptide sets for detection of antiviral CD8 and CD4 T cell responses

Draenert, Rika; Altfeld, Marcus; Berthou, Christian; Basgoz, Nesli; Corcoran, Colleen; Wurcel, Alysse G.; Stone, David R.; Kalams, Spyros A.; Trocha, Alicja; Addo, Marylyn M.; Goulder, Philip; Walker, Bruce D.

doi:10.1016/s0022-1759(02)00541-0

Cited by 126 publications

(101 citation statements)

References 23 publications

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“…This number likely represents an underestimate of the true frequency of induction of WN-specific T cell responses, because the ELISPOT assay was performed by using pools of 20-mer peptides. Peptide length, distribution of epitopes within a peptide, and the number of peptides in the pools can influence sensitivity of the ELISPOT assay (27,28). The number of WN virus-specific T cells was measured by using only peptides to the E protein; on the other hand, the proliferation assay uses an inactivated antigen made from WN virus-infected cells and con- tains all viral structural and nonstructural proteins.…”

Section: Discussionmentioning

confidence: 99%

A live, attenuated recombinant West Nile virus vaccine

Monath

Liu

Kanesa-thasan

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

208

119

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authors note that Fig. 5 did not include all vaccine study groups. The corrected figure and legend appear below. This error does not affect the conclusions of the article. Confidence intervals for Spearman's rank correlation of log 10 IFN-␥ producing PBMC per million and log10 stimulation index were based on Fisher's transformation. On day 14, the correlation was 0.491 (95% CI, 0.231-0.686); on day 28, the correlation was 0.188 (95% CI: Ϫ0.112 to 0.456).

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Section: Discussionmentioning

confidence: 99%

A live, attenuated recombinant West Nile virus vaccine

Monath

Liu

Kanesa-thasan

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

208

119

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“…While important QA procedures such as careful peptide set design (2,8,9,20), biochemical analysis, and selection of appropriate peptide solvent (18) are standard practice for most laboratories, realworld biological assay screening of synthetic peptide sets is not routinely conducted. The selection of an appropriate sample size for the biological screening of the peptides will be dependent upon the HLA diversity in the population studied.…”

Section: Discussionmentioning

confidence: 99%

Peptide Impurities in Commercial Synthetic Peptides and Their Implications for Vaccine Trial Assessment

Currier

Galley

Wenschuh

et al. 2008

Clin Vaccine Immunol

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The advent of T-cell assay methodologies that are amenable to high throughput coupled with the availability of large libraries of overlapping peptides have revolutionized the fields of vaccine efficacy testing and cellular immune response assessment. Since T-cell assay performance is critically dependent upon the quality and specificity of the stimulating peptides, assurance of high-quality and reliable input peptides is an important aspect of assay validation. Herein, we demonstrate that individual peptides from large human immunodeficiency virus (HIV)-based peptide library sets obtained directly from two independent custom peptide suppliers contained contaminating peptides capable of giving false-positive results, which were consistent with nominal antigen-specific CD8 ؉ T-cell responses. In-depth investigation of the cellular response in terms of responding CD8 ؉ T-cell frequency and human leukocyte antigen (HLA) restriction led to the conclusion that one set of HIV type 1 (HIV-1)-derived peptides was contaminated with a peptide from human cytomegalovirus (HCMV), which is commonly used in cellular immunology research applications. Analytical characterization of the original stock of the suspect HIV-1 peptide confirmed the presence of ϳ1% by weight of the HCMV peptide. These observations have critical implications for quality assurance (QA) and quality control (QC) of peptides used in clinical trials where cellular immune-based assays are important end-point determinants. We propose a simple schema of biological QA/QC protocols to augment the standard biochemical QA/QC analyses as a means to circumvent this and other problems that can affect cellular immune-based assay outcome and interpretation.The identification and characterization of immunogenic Tcell epitope-containing regions of the proteomes of infectious agents and candidate cancer-and tumor-specific proteins are an essential phase in the rational development of prophylactic vaccines and immunotherapeutic strategies. Recent methodological advances, particularly enzyme-linked immunospot (ELISPOT) assays and cytokine-based flow cytometry (CFC) assays coupled with overlapping pooled peptide technology, give the opportunity for detailed and precise analyses of specific cellular immune responses (1,3,11,19,22,29). As cellular immune response assays proceed from being used primarily as research tools to be being used as tools for clinical evaluation and assessment of end points in different phases of vaccine development, the required standards of quality assurance (QA)/quality control (QC) for these assays rise dramatically (4,10,16,17,21,27,28). While assay techniques and equipment can be standardized by employing standard operating procedures and assay reagents and can be standardized by use of manufacturer-certified assay kits, a critical component of the assay that is not typically subject to standardized QA/QC is the synthetic peptides used for stimulating the T cells. Synthetic peptides are usually specific to the particular application of the assay, d...

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“…Such diverse repertoires have also been observed measuring IFN-␥ in the same patients (data not shown) being characteristic of chronic HIV infection. 33,[46][47][48][49] This diversity of peptide recognition results, among other factors, from HLA-polygenism and -polymorphism in the human population. It requires extensive peptide library testing to obtain comprehensive information on the HIV-specific CD8 ϩ T cell pool in each individual.…”

Section: Trail-producing Cd8 ϩ T Cells In Hiv Infection 1181mentioning

confidence: 99%

The TRAIL of Helpless CD8⁺T Cells in HIV Infection

Küerten

Asaad²,

Schoenberger³

et al. 2008

AIDS Research and Human Retroviruses

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Our understanding of how CD4 ϩ T cells can regulate CD8 ϩ T cell responses in HIV infection is still incomplete. Recent evidence obtained in mice suggests that CD4 ϩ T cell help is required for efficient CD8 ϩ T cellmediated immunity in chronic infection: CD8 ϩ T cells primed in the absence of such help release the TNF-related apoptosis-inducing ligand TRAIL and undergo apoptosis. Using a novel ELISPOT assay, in the present study we show that CD8 ϩ T cells are also a source of the antigen-specific TRAIL response in HIV-infected patients with CD4 ϩ T cell counts below 200. In patients with CD4 ϩ T cell counts above 200 TRAIL was not detectable. Accordingly, antigens to which patients have likely been exposed when CD4 ϩ T cell levels were high (e.g., influenza, CMV, and EBV) did not induce TRAIL. Within the HIV-positive donor population with low CD4 ϩ T cell counts a dissociation of the interferon-␥ (IFN-␥) and TRAIL response to different HIV peptide epitopes was detectable suggesting impaired immunity to antigens that triggered TRAIL in the absence of IFN-␥. Our findings emphasize that "helpless" CD8 ϩ T cells, i.e., cells that have been primed in the absence of CD4 ϩ T cell help, may play a crucial role in HIV infection. A "helpless" phenotype may impair CD8 ϩ T cell control of HIV and other infections and possibly contribute to the depletion of CD4 ϩ T cells via apoptosis. Immunizations and infections in this "helpless" state might result in ineffective CD8 ϩ T cell responses. 1175

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Comparison of overlapping peptide sets for detection of antiviral CD8 and CD4 T cell responses

Cited by 126 publications

References 23 publications

A live, attenuated recombinant West Nile virus vaccine

A live, attenuated recombinant West Nile virus vaccine

Peptide Impurities in Commercial Synthetic Peptides and Their Implications for Vaccine Trial Assessment

The TRAIL of Helpless CD8⁺T Cells in HIV Infection

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Comparison of overlapping peptide sets for detection of antiviral CD8 and CD4 T cell responses

Cited by 126 publications

References 23 publications

A live, attenuated recombinant West Nile virus vaccine

A live, attenuated recombinant West Nile virus vaccine

Peptide Impurities in Commercial Synthetic Peptides and Their Implications for Vaccine Trial Assessment

The TRAIL of Helpless CD8+T Cells in HIV Infection

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The TRAIL of Helpless CD8⁺T Cells in HIV Infection