Human parainfluenza virus-3 can be targeted by rapidly ex vivo expanded T lymphocytes

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• 作者：Lauren P. McLaughlin¹ ; ² ; Haili Lang¹ ; Elizabeth Williams¹ ; Kaylor E. Wright¹ ; Allison Powell¹ ; Conrad R. Cruz³ ; Anamaris M. Colberg-Poley⁴ ; Cecilia Barese¹ ; Patrick J. Hanley¹ ; ² ; Catherine M. Bollard¹ ; ² ; ⁵ ; Michael D. Keller¹ ; ⁵ ; ^{mkeller@cnmc.org" class="auth_mail" title="E-mail the corresponding author}
• 关键词：antiviral therapy ; cytotoxic T-lymphocytes ; human parainfluenza ; immunodeficiency ; immunotherapy
• 刊名：Cytotherapy
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：18
• 期：12
• 页码：1515-1524
• 全文大小：2219 K

文摘

Human parainfluenza virus-3 (HPIV) is a common cause of respiratory infection in immunocompromised patients and currently has no effective therapies. Virus-specific T-cell therapy has been successful for the treatment or prevention of viral infections in immunocompromised patients but requires determination of T-cell antigens on targeted viruses.

Methods

HPIV3-specific T cells were expanded from peripheral blood of healthy donors using a rapid generation protocol targeting four HPIV3 proteins. Immunophenotyping was performed by flow cytometry. Viral specificity was determined by interferon (IFN)-γ ELISpot, intracellular cytokine staining and cytokine measurements from culture supernatants by Luminex assay. Cytotoxic activity was tested by ⁵¹Cr release and CD107a mobilization assays. Virus-specific T cells targeting six viruses were then produced by rapid protocol, and the phenotype of HPIV3-specific T cells was determined by immunomagnetic sorting for IFN-γ-producing cells.

Results

HPIV3-specific T cells were expanded from 13 healthy donors. HPIV3-specific T cells showed a CD4⁺ predominance (mean CD4:CD8 ratio 2.89) and demonstrated specificity for multiple HPIV3 antigens. The expanded T cells were polyfunctional based on cytokine production but only had a minor cytotoxic component. T cells targeting six viruses in a single product similarly showed HPIV3 specificity, with a predominant effector memory phenotype (CD3⁺/CD45RA^–/CCR7^–) in responder cells.

Discussion

HPIV3-specific T cells can be produced using a rapid ex vivo protocol from healthy donors and are predominantly CD4⁺ T cells with Th1 activity. HPIV3 epitopes can also be successfully targeted alongside multiple other viral epitopes in production of six-virus T cells, without loss of HPIV3 specificity. These products may be clinically beneficial to combat HPIV3 infections by adoptive T-cell therapy in immune-compromised patients.