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NK cell defects in X-linked pigmentary reticulate disorder
Petro Starokadomskyy, … , Daniel D. Billadeau, Ezra Burstein
Petro Starokadomskyy, … , Daniel D. Billadeau, Ezra Burstein
Published November 1, 2019
Citation Information: JCI Insight. 2019;4(21):e125688. https://doi.org/10.1172/jci.insight.125688.
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Research Article Infectious disease Inflammation

NK cell defects in X-linked pigmentary reticulate disorder

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Abstract

X-linked reticulate pigmentary disorder (XLPDR, Mendelian Inheritance in Man #301220) is a rare syndrome characterized by recurrent infections and sterile multiorgan inflammation. The syndrome is caused by an intronic mutation in POLA1, the gene encoding the catalytic subunit of DNA polymerase-α (Pol-α), which is responsible for Okazaki fragment synthesis during DNA replication. Reduced POLA1 expression in this condition triggers spontaneous type I interferon expression, which can be linked to the autoinflammatory manifestations of the disease. However, the history of recurrent infections in this syndrome is as yet unexplained. Here we report that patients with XLPDR have reduced NK cell cytotoxic activity and decreased numbers of NK cells, particularly differentiated, stage V, cells (CD3–CD56dim). This phenotype is reminiscent of hypomorphic mutations in MCM4, which encodes a component of the minichromosome maintenance (MCM) helicase complex that is functionally linked to Pol-α during the DNA replication process. We find that POLA1 deficiency leads to MCM4 depletion and that both can impair NK cell natural cytotoxicity and show that this is due to a defect in lytic granule polarization. Altogether, our study provides mechanistic connections between Pol-α and the MCM complex and demonstrates their relevance in NK cell function.

Authors

Petro Starokadomskyy, Katelynn M. Wilton, Konrad Krzewski, Adam Lopez, Luis Sifuentes-Dominguez, Brittany Overlee, Qing Chen, Ann Ray, Aleksandra Gil-Krzewska, Mary Peterson, Lisa N. Kinch, Luis Rohena, Eyal Grunebaum, Andrew R. Zinn, Nick V. Grishin, Daniel D. Billadeau, Ezra Burstein

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Figure 1

NK cell direct cytotoxicity is affected in XLPDR patients.

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NK cell direct cytotoxicity is affected in XLPDR patients.
(A) Flow cyto...
(A) Flow cytometry quantification of NK cells per milliliter in peripheral blood of XLPDR patients (P1–P5) and unaffected individuals (UA4–UA11). Horizontal bars represent the mean; error bars represent the SD. *P < 0.015, Student’s 2-tailed t test. Data are the aggregate from up to 3 independent measurements. (B) Flow cytometry quantification of NK cells in peripheral blood as a percentage of total lymphocytes. P1–P5 and UA1–UA12 are represented. Horizontal bars represent the mean; error bars represent the SD. *P < 0.0005, Student’s 2-tailed t test. Data are the aggregate of 7 independent measurements spanning up to 8 years. (C) NK cell direct cytotoxicity against 721.221 target cells was assessed using NK cells from unaffected controls (UA1, UA2, UA13) or XLDPR patients (P1 and P2), using effector/target (E/T) ratios of 1 and 5. Bars represent the mean; error bars represent the SEM. *P < 0.0065, and **P < 0.0001 by 2-way ANOVA. Data are the aggregate of 2 independent experiments. (D) NK cell direct cytotoxicity against the 721.221 target cell line was determined over the indicated E/T ratios. Primary NK cells obtained from 5 healthy donors (UA14–UA18) were subjected to POLA1 silencing using 2 siRNA duplexes or a pool of them. The data are representative of 2 independent experiments; error bars represent the SEM. *P < 0.0001 by 2-factor ANOVA comparing siPOLA1 samples against controls. Data are the aggregate of 5 independent experiments. (E) Same as D but using the tumor-derived NK cell line NK92mi. The data are representative of 3 independent experiments; error bars represent the SEM. *P < 0.0001 by 2-way ANOVA comparing siPOLA1 mix against control samples. Data are representative of 2 independent experiments. (F) Antibody-dependent cell cytotoxicity (ADCC) of NK cells from unaffected controls (UA8–UA10) and XLPDR patients (P1 and P2). SKOV-3 ovarian cancer cells treated with anti–human epidermal growth factor receptor 2 (anti-HER2) antibodies were used as the target cell line; cells incubated with control antibody (IgG) were used as a negative control. Bars represent the mean; error bars represent the SEM. NS, nonsignificant; P > 0.1 by 2-factor ANOVA comparing UA to XLPDR groups. Data are from a single experiment.

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