Defining blood processing parameters for optimal detection of cryopreserved antigen-specific responses for HIV vaccine trials

Bull, Marta E.; Lee, Dong Hoon; Stucky, Jason; Chiu, Ya Lin; Rubin, Abbe; Horton, Helen; McElrath, M. Juliana

doi:10.1016/j.jim.2007.02.003

Cited by 211 publications

(222 citation statements)

References 26 publications

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“…The loss of responses cannot be attributed to cryopreservation technique or to the shipping of samples, as suggested in previous studies (Disis et al, 2006;Bull et al, 2007;Kierstead et al, 2007), as similar losses were measured in samples processed and frozen at three independent laboratories. These changes occurred despite the observation that cell viability was generally maintained at acceptable levels of 81-95% (Table 2).…”

Section: Discussionsupporting

confidence: 76%

“…The reason for these inconsistent observations has not been well defined. Two recent studies show that the time elapsed between phlebotomy and cryopreservation can result in lower functional responses (Bull et al, 2007;Kierstead et al, 2007). Other factors may include the quality of the cryopreservation and storing procedures (which are generally not standardized), as well as the duration of cryopreservation.…”

Section: Introductionmentioning

confidence: 99%

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Loss of T cell responses following long-term cryopreservation

Owen

Sinclair

Emu

et al. 2007

Journal of Immunological Methods

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Although cryopreservation of peripheral blood mononuclear cells (PBMC) is a commonly used technique, the degree to which it affects subsequent functional studies has not been well defined.Here we demonstrate that long-term cryopreservation has detrimental effects on T cell IFN-γ responses in human immunodeficiency virus (HIV) infected individuals. Long-term cryopreservation caused marked decreases in CD4 + T cell responses to whole proteins (HIV p55 and cytomegalovirus (CMV) lysate) and HIV peptides, and more limited decreases in CD8 + T cell responses to whole proteins. These losses were more apparent in cells stored for greater than one year compared to less than six months. CD8 + T cell responses to peptides and peptide pools were well preserved. Loss of both CD4 + and CD8 + T cell responses to CMV peptide pools were minimal in HIV-negative individuals. Addition of exogenous antigen presenting cells (APC) did not restore CD4 + T cell responses to peptide stimulation and partially restored T cell IFN-γ responses to p55 protein.Overnight resting of thawed cells did not restore T cell IFN-γ responses to peptide or whole protein stimulation. A selective loss of phenotypically defined effector cells did not explain the decrement of responses, although cryopreservation did increase CD4 + T cell apoptosis, possibly contributing to the loss of responses. These data suggest that the impact of cryopreservation should be carefully considered in future vaccine and pathogenesis studies. In HIV-infected individuals short-term cryopreservation may be acceptable for measuring CD4 + and CD8 + T cell responses. Long-term cryopreservation, however, may lead to the loss of CD4 + T cell responses and mild skewing of T cell phenotypic marker expression.

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Section: Discussionsupporting

confidence: 76%

Section: Introductionmentioning

confidence: 99%

Loss of T cell responses following long-term cryopreservation

Owen

Sinclair

Emu

et al. 2007

Journal of Immunological Methods

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“…PBMC were isolated from blood as previously described (57). RNA was extracted from PBMC using the RNeasy Protect Cell protocol (Qiagen).…”

Section: Methodsmentioning

confidence: 99%

Merck Ad5/HIV induces broad innate immune activation that predicts CD8 ⁺ T-cell responses but is attenuated by preexisting Ad5 immunity

Zak

Andersen‐Nissen²,

Peterson³

et al. 2012

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

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To better understand how innate immune responses to vaccination can lead to lasting protective immunity, we used a systems approach to define immune signatures in humans over 1 wk following MRKAd5/HIV vaccination that predicted subsequent HIV-specific T-cell responses. Within 24 h, striking increases in peripheral blood mononuclear cell gene expression associated with inflammation, IFN response, and myeloid cell trafficking occurred, and lymphocyte-specific transcripts decreased. These alterations were corroborated by marked serum inflammatory cytokine elevations and egress of circulating lymphocytes. Responses of vaccinees with preexisting adenovirus serotype 5 (Ad5) neutralizing antibodies were strongly attenuated, suggesting that enhanced HIV acquisition in Ad5-seropositive subgroups in the Step Study may relate to the lack of appropriate innate activation rather than to increased systemic immune activation. Importantly, patterns of chemoattractant cytokine responses at 24 h and alterations in 209 peripheral blood mononuclear cell transcripts at 72 h were predictive of subsequent induction and magnitude of HIV-specific CD8 + T-cell responses. This systems approach provides a framework to compare innate responses induced by vectors, as shown here by contrasting the more rapid, robust response to MRKAd5/HIV with that to yellow fever vaccine. When applied iteratively, the findings may permit selection of HIV vaccine candidates eliciting innate immune response profiles more likely to drive HIV protective immunity.

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“…Serum for humoral assays was obtained from serum-separating tubes (SSTs) and frozen at -80°C until use. Peripheral blood mononuclear cells (PBMCs) for cellular assays were isolated and cryopreserved from heparin-anticoagulated whole blood within 6 hours of venipuncture, as described previously (35). PBMCs were thawed and cultured overnight at 37°C, 5% CO 2 in R10 (RPMI 1640 [Gibco], 10% FBS [Gemini Bioproducts], 2 mM L-glutamine [Gibco], 100 μg/ml streptomycin sulfate [Gibco], and 100 U/ml penicillin G [Gibco]).…”

Section: Functionality Of Responses Is Similar Across All Tested Regimentioning

confidence: 99%

HIV-specific humoral responses benefit from stronger prime in phase Ib clinical trial

Bart¹,

Huang²,

Karuna³

et al. 2014

J. Clin. Invest.

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BACKGROUND. Vector prime-boost immunization strategies induce strong cellular and humoral immune responses. We examined the priming dose and administration order of heterologous vectors in HIV Vaccine Trials Network 078 (HVTN 078), a randomized, double-blind phase Ib clinical trial to evaluate the safety and immunogenicity of heterologous prime-boost regimens, with a New York vaccinia HIV clade B (NYVAC-B) vaccine and a recombinant adenovirus 5-vectored (rAd5-vectored) vaccine. METHODS.NYVAC-B included HIV-1 clade B Gag-Pol-Nef and gp120, while rAd5 included HIV-1 clade B Gag-Pol and clades A, B, and C gp140. Eighty Ad5-seronegative subjects were randomized to receive 2 × NYVAC-B followed by 1 × 10 10 PFU rAd5 (NYVAC/Ad5 hi ); 1 × 10 8 PFU rAd5 followed by 2 × NYVAC-B (Ad5 lo /NYVAC); 1 × 10 9 PFU rAd5 followed by 2 × NYVAC-B (Ad5 med /NYVAC); 1 × 10 10 PFU rAd5 followed by 2 × NYVAC-B (Ad5 hi /NYVAC); or placebo. Immune responses were assessed 2 weeks after the final vaccination. Intracellular cytokine staining measured T cells producing IFN-γ and/or IL-2; cross-clade and epitope-specific binding antibodies were determined; and neutralizing antibodies (nAbs) were assessed with 6 tier 1 viruses. RESULTS. CD4+ T cell response rates ranged from 42.9% to 93.3%. NYVAC/Ad5 hi response rates (P ≤ 0.01) and magnitudes (P ≤ 0.03) were significantly lower than those of other groups. CD8+ T cell response rates ranged from 65.5% to 85.7%. NYVAC/Ad5 hi magnitudes were significantly lower than those of other groups (P ≤ 0.04). IgG response rates to the group M consensus gp140 were 89.7% for NYVAC/Ad5 hi and 21.4%, 84.6%, and 100% for Ad5 lo /NYVAC, Ad5 med /NYVAC, and Ad5 hi /NYVAC, respectively, and were similar for other vaccine proteins. Overall nAb responses were low, but aggregate responses appeared stronger for Ad5 med /NYVAC and Ad5 hi /NYVAC than for NYVAC/Ad5 hi . CONCLUSIONS. rAd5 prime followed by NYVAC boost is superior to the reverse regimen for both vaccine-induced cellular and humoral immune responses. Higher Ad5 priming doses significantly increased binding and nAbs. These data provide a basis for optimizing the design of future clinical trials testing vector-based heterologous prime-boost strategies.TRIAL REGISTRATION. ClinicalTrials.gov NCT00961883.FUNDING. NIAID, NIH UM1AI068618, AI068635, AI068614, and AI069443.

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Defining blood processing parameters for optimal detection of cryopreserved antigen-specific responses for HIV vaccine trials

Cited by 211 publications

References 26 publications

Loss of T cell responses following long-term cryopreservation

Loss of T cell responses following long-term cryopreservation

Merck Ad5/HIV induces broad innate immune activation that predicts CD8 ⁺ T-cell responses but is attenuated by preexisting Ad5 immunity

HIV-specific humoral responses benefit from stronger prime in phase Ib clinical trial

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Defining blood processing parameters for optimal detection of cryopreserved antigen-specific responses for HIV vaccine trials

Cited by 211 publications

References 26 publications

Loss of T cell responses following long-term cryopreservation

Loss of T cell responses following long-term cryopreservation

Merck Ad5/HIV induces broad innate immune activation that predicts CD8 + T-cell responses but is attenuated by preexisting Ad5 immunity

HIV-specific humoral responses benefit from stronger prime in phase Ib clinical trial

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Product

Resources

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Merck Ad5/HIV induces broad innate immune activation that predicts CD8 ⁺ T-cell responses but is attenuated by preexisting Ad5 immunity