Effect of Cryopreservation on Measurement of Cytomegalovirus-Specific Cellular Immune Responses in HIV-Infected Patients

Weinberg, Adriana; Wohl, David A.; Brown, Darby; Pott, Gregory B.; Zhang, Li; Ray, Michelle; Horst, Charles van der

doi:10.1097/00126334-200010010-00004

Cited by 17 publications

(9 citation statements)

References 12 publications

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“…Lymphocyte subsets and proliferative responses were measured in parallel and were compared with those of age-specific control subjects. VZV-specific lymphocyte proliferation was performed as described elsewhere [17]. NK-cell cytolytic activity was assessed by a standard K562 target-cell-lysis assay measuring [ 51 Cr] release [18].…”

Section: Patient Materials and Methodsmentioning

confidence: 99%

Disseminated Varicella Infection Due to the Vaccine Strain of Varicella‐Zoster Virus, in a Patient with a Novel Deficiency in Natural Killer T Cells

Levy¹,

Orange²,

Hibberd³

et al. 2003

J INFECT DIS

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164

107

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An 11-year-old girl presented with a papulovesicular rash and severe respiratory distress 5 weeks after receiving varicella vaccine. Restriction fragment length-polymorphism analysis of virus isolated from an endotracheal-tube aspirate and from bronchoalveolar lavage revealed that this patient's illness was due to the Oka vaccine strain of varicella. An extensive immunologic analysis failed to identify a known diagnostic entity to explain her susceptibility to this attenuated vaccine strain. Analysis of her lymphocytes on separate occasions, months after recovery from her illness, revealed a profound deficiency of natural killer T (NKT) cells and of NKT-cell activity, suggesting that NKT cells contribute to host defense against varicella virus.

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Section: Patient Materials and Methodsmentioning

confidence: 99%

Disseminated Varicella Infection Due to the Vaccine Strain of Varicella‐Zoster Virus, in a Patient with a Novel Deficiency in Natural Killer T Cells

Levy¹,

Orange²,

Hibberd³

et al. 2003

J INFECT DIS

Self Cite

164

107

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“…The lymphocyte proliferation assay (LPA) was performed as previously described (36). PBMC were resuspended in RPMI 1640 (Gibco) with glutaminecontaining 10% human AB serum (Nabi) and 1% antibiotics (Gibco) (stimulation medium).…”

Section: Patientsmentioning

confidence: 99%

“…After 6 h of incubation at 37°C in a CO 2 incubator, brefeldin A (Sigma) was added to a final concentration of 10 g/ml. Cells were further incubated for 16 h at 37°C in a CO 2 incubator and then fixed and stained for CD69, gamma interferon (IFN-␥), CD4, and isotype controls (36). At least 10,000 cells were analyzed in each assay by use of a Coulter XL flow cytometer.…”

Section: Patientsmentioning

confidence: 99%

Immunity to Human Immunodeficiency Virus (HIV) in Children with Chronic HIV Infection Receiving Highly Active Antiretroviral Therapy

Weinberg

Pott

2003

Clin Vaccine Immunol

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Our objective was to describe the CD4-mediated human immunodeficiency virus (HIV)-specific cell-mediated immunity (CMI) and its virologic and immunologic correlates in children with chronic HIV infection on highly active antiretroviral therapy (HAART). Twelve HIV-infected children on stable antiretroviral therapy with a median level of CD4؉ lymphocytes (CD4%) of 25.5% and a median viral load (VL) of 786 HIV RNA copies/ml were enrolled in this study. Nine of these children were also cytomegalovirus (CMV) seropositive. Blood mononuclear cells, stimulated with HIV and CMV antigens, were used to measure lymphocyte proliferation and to enumerate gamma interferon (IFN-␥)-producing CD4 ؉ cells. HIV CMI and CMV CMI were detected in similar proportions of patients and correlated with each other, although the HIV responses were less robust. HIV lymphocyte proliferation significantly increased with lower HIV VL and showed a trend to increase with higher CD4% and longer time on HAART. The in vitro IFN-␥ response to HIV or CMV was not affected by CD4%, VL, or HAART. Pediatric patients with established HIV infection on HAART frequently exhibit HIV CMI despite undetectable HIV replication. We concluded that the association between HIV CMI and CMV CMI indicates that the same factors govern responsiveness to either antigen. Human immunodeficiency virus (HIV) preferentially destroys CD4ϩ T lymphocytes and interferes with the functioning of the immune system, weakening defenses against infectious agents. Highly active antiretroviral therapy (HAART) restores CD4 ϩ cell numbers and greatly decreases the incidence of opportunistic infections, indicating that significant immune recovery occurs in treated patients (17,23,28). Furthermore, individuals who previously met AIDS diagnostic criteria recover cytomegalovirus (CMV)-, Mycobacterium avium-, Mycobacterium intracellulare-, and Candida-specific cell-mediated immunity (CMI) while on HAART (8,15,18).AIDS-associated impairment of the immune system includes the inability of the host to mount a protective response against HIV (21, 27). Although HIV-specific CD8-mediated cytotoxicity as detected in HIV-infected patients plays a protective role against the progression of infection (6, 13, 25), CD4-mediated immunity has been difficult to identify. Long-term nonprogressors and patients treated soon after primary infection display HIV-specific CD4-dependent lymphocyte proliferation (1, 20), but most chronically infected adults, including HAART recipients, do not (24, 29).The objective of this study was to compare the HIV-and CMV-specific CD4-mediated responses of HIV-infected children on HAART by using two recently developed and highly sensitive immunological assays, the enzyme-linked immunospot (ELISPOT) assay (32) and intracellular cytokine flow enumeration (ICCK) (34), coupled with an inactivated whole-HIV antigen preparation (3, 26).(Part of this study was presented at the 9th Conference on Retroviruses and Opportunistic Infections, 2002.) MATERIALS AND METHODS Patients.The study enrolled...

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“…The effect of cryopreservation on T cell responses has been more extensively studied, but results have been mixed. Although cryopreservation of PBMCs can alter cell surface markers, [10][11][12] cytokine production, 11,13 and cell proliferation, 14 such discrepancies can be minimized when an optimized protocol is used. 12,[15][16][17] Preservation of cell function has been shown to be improved if PBMCs are frozen within 12 h of blood collection 17 and temperature fluctuations are minimized during storage.…”

Section: Introductionmentioning

confidence: 99%

Effect of cryopreservation of peripheral blood mononuclear cells (PBMCs) on the variability of an antigen-specific memory B cell ELISpot

Trück

Mitchell²,

Thompson

et al. 2014

Human Vaccines & Immunotherapeutics

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Effect of Cryopreservation on Measurement of Cytomegalovirus-Specific Cellular Immune Responses in HIV-Infected Patients

Cited by 17 publications

References 12 publications

Disseminated Varicella Infection Due to the Vaccine Strain of Varicella‐Zoster Virus, in a Patient with a Novel Deficiency in Natural Killer T Cells

Disseminated Varicella Infection Due to the Vaccine Strain of Varicella‐Zoster Virus, in a Patient with a Novel Deficiency in Natural Killer T Cells

Immunity to Human Immunodeficiency Virus (HIV) in Children with Chronic HIV Infection Receiving Highly Active Antiretroviral Therapy

Effect of cryopreservation of peripheral blood mononuclear cells (PBMCs) on the variability of an antigen-specific memory B cell ELISpot

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