Selective ex vivo expansion of cytomegalovirus‐specific CD4+ and CD8+ T lymphocytes using dendritic cells pulsed with a human leucocyte antigen A*0201‐restricted peptide

Vannucchi, Alessandro M.; Glinz, S; Bosi, Alberto; Caporale, Roberto; Rossi-Ferrini, P

doi:10.1046/j.1365-2141.2001.02777.x

Cited by 22 publications

(11 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Current approaches are time consuming, require leukapheresis of donor for the generation of virus-specific T cells, use replicative competent virus, which is prohibited under current GMP regulations, or isolate CD4 ϩ or CD8 ϩ virus-specific T cells for adoptive transfer. [17][18][19][20][21][22] This study was conducted to circumvent limitations in the generation of virus-specific T-cell lines for adoptive transfer into allogeneic SCT recipients without impairing the functionality of the generated T-cell lines. To facilitate the enrichment of virusspecific T cells, the novel selection approach of the IFN-␥ secretion assay was applied to current GMP regulations, and a methodology was developed for the rapid enrichment and expansion of combined CD4 ϩ and CD8 ϩ CMV-specific T cells.…”

Section: Discussionmentioning

confidence: 99%

“…Ex vivo induction of CMV-specific T cells using CMV-infected autologous fibroblasts, EBV-infected B cells, or antigen-presenting cells (APCs) transduced with genes of interest as stimulator cells 13,[17][18][19] is an effective approach, but the potential biohazard resulting from the presence of live viruses may not be applicable to current good manufacturing practice (GMP) standards. Generating CMV-specific T cells using CMV peptidepulsed dendritic cells (DCs) 20,21 or CMV antigen-pulsed DCs 17,22 or using genetically modified APCs 18,19 is also an effective approach, but each is labor intensive and requires several weeks for the generation of a clinical product. Thus, alternative strategies are warranted to avoid the application of replicative virus during the stimulation procedure and to reduce the time required to generate sufficient CMV-specific T cells for adoptive transfer.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Rapid generation of combined CMV-specific CD4+ and CD8+ T-cell lines for adoptive transfer into recipients of allogeneic stem cell transplants

Rauser

Einsele

Sinzger

et al. 2004

Blood

156

123

View full text Add to dashboard Cite

Adoptive transfer of cytomegalovirus (CMV)-specific T cells can restore longlasting, virus-specific immunity and clear CMV viremia in recipients of allogeneic stem cell transplants if CD4 ؉ and CD8 ؉ CMV-specific T cells are detected in the recipient after transfer. Current protocols for generating virus-specific T cells use live virus, require leukapheresis of the donor, and are time consuming. To circumvent these limitations, a clinicalscale protocol was developed to generate CMV-specific T cells by using autologous cellular and serum components derived from a single 500-mL blood draw. CMVspecific T cells were stimulated simultaneously with CMV-specific major histocompatibility complex class I (MHC I)-restricted peptides and CMV antigen. Activated T cells were isolated with the interferon-␥ (IFN-␥) secretion assay and expanded for 10 days. In 8 randomly selected, CMV-seropositive donors, 1.34 ؋ 10 8 combined CD4 ؉ and CD8 ؉ CMV-specific T cells, on average, were generated, as determined by antigentriggered IFN-␥ production. CMV-infected fibroblasts were efficiently lysed by the generated T cells, and CMV-specific CD4 ؉ and CD8 ؉ T cells expanded if they were stimulated with natural processed antigen. On the other hand, CD4 ؉ and CD8 ؉ T cell-mediated alloreactivity of generated CMV-specific T-cell lines was reduced compared with that of the starting population. In conclusion, the culture system developed allowed the rapid generation of allodepleted, highly enriched, combined CD4 ؉ and CD8 ؉ CMV-specific T cells under conditions mimicking good manufacturing practice.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Rapid generation of combined CMV-specific CD4+ and CD8+ T-cell lines for adoptive transfer into recipients of allogeneic stem cell transplants

Rauser

Einsele

Sinzger

et al. 2004

Blood

156

123

View full text Add to dashboard Cite

show abstract

“…This could account at least in part for the lower anti-HHV-8 T-cell responses detected in healthy, HHV-8-seropositive individuals, [2][3][4] as compared to T-cell responses to EBV and CMV peptides in healthy, EBV-and CMV-seropositive persons. [36][37][38][39] Although the precise function of HHV-8 gB is unknown, a recent report shows that it is part of the virion and infected cell membrane, 49 in contrast to gB of EBV and MHV-68. 50,51 Indeed, gB may be important for HHV-8 infectivity and pathogenesis in that it binds to cell surface heparan sulfate molecules.…”

Section: Discussionmentioning

confidence: 99%

“…We and others have previously shown that human blood monocytederived DCs loaded with HIV-1, 35 EBV, 36,37 and human cytomegalovirus (CMV) 38,39 peptides are potent inducers of antiviral CD8 ϩ T-cell responses in PBMCs of individuals seropositive for these viruses. Therefore, we determined if autologous DCs loaded with gB 492-500 peptide were able to induce peptide-specific T-cell reactivity as compared to the conventional method of stimulation with peptide alone.…”

Section: T-cell Response To Gb 492-500 In Hla A*0201 Hhv-8 Seropositimentioning

confidence: 99%

Identification of an HLA A*0201–restricted CD8+T-cell epitope for the glycoprotein B homolog of human herpesvirus 8

Wang

Huang

Rappocciolo

et al. 2002

Blood

View full text Add to dashboard Cite

IntroductionHuman herpesvirus 8 (HHV-8; also termed Kaposi sarcomaassociated herpesvirus) is a gammaherpesvirus that is considered to be the causative agent of Kaposi sarcoma (KS), body cavity B-cell lymphoma, and multicentric Castleman disease. 1 T-cell immunity is thought to play an important role in control of HHV-8 infection and these associated diseases. [2][3][4][5] In this regard, we 2,3 and others 4 have demonstrated that major histocompatibility complex (MHC) class I-restricted, CD8 ϩ cytotoxic T lymphocytes (CTLs), and interferon ␥ (IFN-␥) responses are detectable in peripheral blood mononuclear cells (PBMCs) of HHV-8 ϩ individuals using recombinant vaccinia viruses expressing HHV-8 lytic and latent cycle proteins. These anti-HHV-8 T-cell responses are lower during human immunodeficiency virus type 1 (HIV-1) infection and may be involved in prevention of KS and other HHV-8-associated diseases.Support for the potential role of CD8 ϩ T-cell immunity in control of HHV-8 infection comes from several studies on immune control of acute and persistent infections with 2 other gammaherpesviruses, specifically, Epstein-Barr virus (EBV) and murine herpesvirus 68 (MHV-68). 6-10 Although EBV latency proteins can induce strong CD8 ϩ T-cell responses, 11 lytic cycle proteins are considered as major CD8 ϩ T-cell targets in both acute and persistent EBV infections. 12 Immunodominant epitopes restricted to different HLA class I alleles, which are important for our understanding of viral immunopathogenesis and development of appropriate vaccines, 13 have been determined for peptides derived from EBV structural proteins gp350 and gp85, and immediate-early (IE) and early proteins such as BZLF1, BRLF1, and BMLF1. [6][7][8][9] Compared to latency proteins, the frequency of CD8 ϩ T lymphocytes specific for peptides derived from lytic cycle proteins is higher in PBMCs of healthy virus carriers. 14 CD8 ϩ CTLs have also been demonstrated to secrete IFN-␥ in response to EBV lytic cycle antigens in seropositive individuals. 15 During primary EBV infectious mononucleosis, up to 60% of CD8 ϩ T cells circulating in blood are EBV-specific. 16 Furthermore, by use of a peptide-tetramer complex, Tan et al 17 have directly visualized from 0.5% to 6.6% of CD8 ϩ T blood cells in virus carriers that are specific for a single BMLF-1 epitope.In MHV-68 infection, lytic cycle proteins also induce CD8 ϩ T cell responses. 10,18,19 T cells specific for peptides derived from lytic cycle proteins dominate the acute phase of MHV-68 infection. 10,19 Moreover, vaccination with immunodominant CD8 ϩ T-cell epitopes derived from lytic cycle proteins of MHV-68 significantly reduces viral titers and the level of infected cells during acute infection. 20 We have tested CD8 ϩ T-cell reactivity to peptides derived from 5 HHV-8 lytic cycle proteins based on a prediction model for the HLA A*0201 motif in HHV-8-infected subjects to determine immunodominant epitopes to the virus. We chose HLA A*0201 because it is prevalent in a large percentage of the white populat...

show abstract

“…This further confirms that peptideinduced CD8 T-cell lines recognize CMV antigens, as shown also by other groups. 19,[21][22][23] Also in this set of donors specificity was confirmed and frequency of specific T cells was remarkable. Nevertheless, we were unable to reduce phenotype variability, suggesting that more technologic improvements must be pursued.…”

Section: Functional Assays On Expanded T-cell Lines Obtained From Thementioning

confidence: 65%

Positive Selection and Expansion of Cytomegalovirus-specific CD4 and CD8 T Cells in Sealed Systems

Pira¹,

Ivaldi

Tripodi

et al. 2008

Journal of Immunotherapy

View full text Add to dashboard Cite

Administration of pathogen-specific T-cell lines can reconstitute the cellular immune function of immunocompromised patients. Selection and expansion of specific T cells for reinfusion pose unique challenges owing to the fact that good manufacturing procedures must be implemented. Cytokine secretion-based methods can identify and select specific T cells. We showed here that it is possible to combine this method with procedures for cell handling performed in a sealed, unbreached system from start to end. Peripheral blood mononuclear cells, obtained from blood samples and processed in a sealed system, were stimulated in Teflon bags with a library of selected CD4 and CD8 peptides derived from the immunodominant cytomegalovirus protein pp65. The stimulated T cells were labeled with reagents for interferon-gamma surface capture and selected on a magnetic column using a sealed system connected to the Teflon bags. Elution and final expansion were also performed with an unbreached protocol with preservation of sterility even if the steps were run on the bench top. Expanded cells exhibited the appropriate functions. The use of this unbreached procedure proves that safety of cellular products generated in a good manufacturing procedures facility can be further improved. Similar sealed protocols can also be applied for T-cell therapies directed against tumor antigens.

show abstract

Selective ex vivo expansion of cytomegalovirus‐specific CD4⁺ and CD8⁺ T lymphocytes using dendritic cells pulsed with a human leucocyte antigen A*0201‐restricted peptide

Cited by 22 publications

References 15 publications

Rapid generation of combined CMV-specific CD4+ and CD8+ T-cell lines for adoptive transfer into recipients of allogeneic stem cell transplants

Rapid generation of combined CMV-specific CD4+ and CD8+ T-cell lines for adoptive transfer into recipients of allogeneic stem cell transplants

Identification of an HLA A*0201–restricted CD8+T-cell epitope for the glycoprotein B homolog of human herpesvirus 8

Positive Selection and Expansion of Cytomegalovirus-specific CD4 and CD8 T Cells in Sealed Systems

Contact Info

Product

Resources

About