NK Cells from RAG- or DCLRE1C-Deficient Patients Inhibit HCMV.

Zeguang Wu,Narmadha Subramanian,Johannes Van Der Merwe,Thomas Mertens,Kerstin Laib Sampaio,Eva-Maria Jacobsen,Manfred Hönig

doi:10.3390/microorganisms7110546

Zeguang Wu, Narmadha Subramanian + Show 5 more

Open Access

https://doi.org/10.3390/microorganisms7110546

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Abstract

The recombination-activating genes (RAGs) and the DNA cross-link repair 1C gene (DCLRE1C) encode the enzymes RAG1, RAG2 and Artemis. They are critical components of the V(D)J recombination machinery. V(D)J recombination is well known as a prerequisite for the development and antigen diversity of T and B cells. New findings suggested that RAG deficiency impacts the cellular fitness and function of murine NK cells. It is not known whether NK cells from severe combined immunodeficiency (SCID) patients with defective RAGs or DCLRE1C (RAGs−/DCLRE1C−-NK) are active against virus infections. Here, we evaluated the anti-HCMV activity of RAGs−/DCLRE1C−-NK cells. NK cells from six SCID patients were functional in inhibiting HCMV transmission between cells in vitro. We also investigated the expansion of HCMV-induced NK cell subset in the RAG- or DCLRE1C-deficient patients. A dynamic expansion of NKG2C+ NK cells in one RAG-2-deficient patient was observed post HCMV acute infection. Our study firstly reveals the antiviral activity of human RAGs−/ DCLRE1C−-NK cells.

Highlights

HCMV is a ubiquitous herpes virus that persists in the host
By using our HCMV transmission inhibition assay [11], we firstly investigated whether recombination-activating genes (RAGs)−/DNA cross-link repair 1C gene (DCLRE1C)−-NK cells can inhibit the HCMV transmission in cell cultures
We found that T cells and NK cells from healthy donor Peripheral blood mononuclear cells (PBMCs) are effectors in inhibiting HCMV transmission, whereas B cells are not involved

Summary

Introduction

HCMV is a ubiquitous herpes virus that persists in the host. Control of HCMV requires a continuous immune surveillance, including innate and adaptive immunity. The key components involved are RAG1 and 2, terminal deoxynucleotidyl transferase, Artemis nuclease, DNA-dependent protein kinase, X-ray repair cross-complementing protein 4, DNA ligase IV, non-homologous end-joining factor 1 and DNA polymerases λ and μ [2,3] Mutations in these genes result in V(D)J recombination deficiency [4,5]. NK cells are generally considered as effectors of innate immunity because they lack antigen-specific cell surface receptors [8] Their activities are regulated by activating and inhibitory receptors and are supposed to not require RAGs and DCLRE1C. The patient had normal immune functions, except for an extreme deficiency of NK cells This case report suggested the importance of NK cells in controlling herpesviruses infections including active HCMV infection [10]. The anti-HCMV role of NK cells from SCID patients with defective RAGs or DCLRE1C (RAGs−/DCLRE1C−-NK) is unknown

Patients and Cells

Preparation of Viral Stocks and Focal Expansion Assay

Flow Cytometry and IFN-γ Detection

Results

Discussion