Strategic use of an adenoviral vector for rapid and efficient ex vivo-generation of cytomegalovirus pp65-reactive cytolytic and helper T cells

• Cytomegalovirus (CMV) reactivation can cause severe complications for transplant patients. Such patients can be protected against CMV-associated diseases through reconstitution of donor-derived CMV-reactive cytolytic and helper T cells. We have developed a strategic protocol for efficient simultaneous generation of CMV-reactive CD8 + and CD4 + T cells ex vivo . The protocol uses peripheral blood lymphocytes (PBLs), antigen-modified mature dendritic cells (DCs) generated in only 3 days and an adenoviral vector encoding the CMV pp65 antigen (Adpp65) both as an endogenous and exogenous source of antigen. PBLs stimulated once with Adpp65-transduced DCs (endogenously expressed pp65) resulted in preferential activation and expansion of pp65-specific CD8 + T cells while PBLs stimulated with DCs pulsed with cell lysate from Adpp65-transduced autologous monocytes (exogenously expressed pp65) yielded pp65-specific CD4 + T cells. Stimulation with double-modified DCs efficiently activated and expanded cytolytic and helper T cells simultaneously. The frequency of T cells producing interferon (IFN)-γ in response to pp65 increased after one stimulation on average 9.6-fold to 4.3% for CD8 + T cells and 25.8-fold to 6.5% for CD4 + T cells. This implies that sufficient number of pp65-specific cytolytic and helper T cells for adoptive transfer may be obtained in only two weeks.

Ämnesord

Medical and Health Sciences  (hsv)

Basic Medicine  (hsv)

Microbiology in the medical area  (hsv)

Medicin och hälsovetenskap  (hsv)

Medicinska grundvetenskaper  (hsv)

Mikrobiologi inom det medicinska området  (hsv)

Medical and Health Sciences  (hsv)

Basic Medicine  (hsv)

Immunology in the medical area  (hsv)

Medicin och hälsovetenskap  (hsv)

Medicinska grundvetenskaper  (hsv)

Immunologi inom det medicinska området  (hsv)

MEDICINE  (svep)

Microbiology, immunology, infectious diseases  (svep)

MEDICIN  (svep)

Mikrobiologi, immunologi, infektionssjukdomar  (svep)

MEDICINE  (svep)

Microbiology, immunology, infectious diseases  (svep)

Immunology  (svep)

Clinical immunology  (svep)

MEDICIN  (svep)

Mikrobiologi, immunologi, infektionssjukdomar  (svep)

Immunologi  (svep)

Klinisk immunologi  (svep)

Klinisk immunologi  (uu)

Clinical Immunology  (uu)

Nyckelord

Dendritic cells

T lymphocytes

Cytomegalovirus

Adenoviral vector

pp65