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



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 by51Cr 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.



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.



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.
Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA; Division of Blood and Marrow Transplantation, Children's National Medical Center, Washington, DC, USA.
Lauren P
LP
McLaughlin
Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA.
Haili
H
Lang
Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA.
Elizabeth
E
Williams
Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA.
Kaylor E
KE
Wright
Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA.
Allison
A
Powell
Sheikh Zayed Institute, Children's National Medical Center, Washington, DC, USA.
Conrad R
CR
Cruz
Center for Genetic Medicine Research, Children's National Medical Center, Washington, DC, USA.
Anamaris M
AM
Colberg-Poley
Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA.
Cecilia
C
Barese
Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA; Division of Blood and Marrow Transplantation, Children's National Medical Center, Washington, DC, USA.
Patrick J
PJ
Hanley
Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA; Division of Blood and Marrow Transplantation, Children's National Medical Center, Washington, DC, USA; Division of Allergy and Immunology, Children's National Medical Center, Washington, DC, USA.
Catherine M
CM
Bollard
Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA; Division of Allergy and Immunology, Children's National Medical Center, Washington, DC, USA. Electronic address: mkeller@cnmc.org.
Michael D
MD
Keller
D000956:Antigens, Viral / Q000276:immunology*; D016516:CD4-CD8 Ratio; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D002478:Cells, Cultured; D005434:Flow Cytometry; D006801:Humans; D016867:Immunocompromised Host; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000276:immunology; D018655:Lymphocyte Count; D010224:Parainfluenza Virus 3, Human / Q000276:immunology*; D010253:Respirovirus Infections / Q000276:immunology / Q000628:therapy*
D000956:Antigens, Viral; D007371:Interferon-gamma
antiviral therapy; cytotoxic T-lymphocytes; human parainfluenza; immunodeficiency; immunotherapy
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