Distinct Immunity Dynamics of Natural Killer Cells in Mild and Moderate COVID-19 Cases During the Omicron Variant Phase

Yukari Nishikawa¹Kosuke Yamaguchi^2*Ma’arif Athok Shofiudin¹Momone Mimura³Miyako Takata³Shu Mihara¹Takeru Kawakami¹Ayumu Doi¹Risa Matsuda¹Hiroyuki Kato¹Ryo Okamoto¹Kengo Mukuda¹Naoki Kinoshita⁴Kensaku Okada¹Tsuyoshi Kitaura¹Masaki Nakamoto¹Hisashi Noma⁵Yusuke Endo⁶Akira Yamasaki²Hiroki Chikumi¹

• ¹Division of Infectious Diseases, School of Medicine, Faculty of Medicine, Tottori University, Yonago, Japan
• ²Department of Multidisciplinary Internal Medicine, Division of Respiratory Medicine and Rheumatology, Faculty of Medicine, Tottori University, Yonago, Japan
• ³Department of Pathobiological Science and Technology, Faculty of Medicine, Tottori University, Yonago, Japan
• ⁴Division of Respiratory Medicine and Rheumatology, Department of Multidisciplinary Internal Medicine, Faculty of Medicine, Tottori University, Yonago, Tottori, Japan
• ⁵Department of Interdisciplinary Statistical Mathematics, The Institute of Statistical Mathematics, Tokyo, Japan
• ⁶Organisation for Research Institute and Promotion, Tottori University, Yonago, Japan

The SARS-CoV-2 Omicron variant is associated with milder COVID-19 symptoms than previous strains. This study analyzed alterations in natural killer (NK) cell-associated immunity dynamics in mild and moderate COVID-19 cases during the Omicron phase of the COVID-19 pandemic.We conducted a retrospective observational cohort study of patients aged ≥16 with confirmed SARS-CoV-2 infection who were hospitalized at Tottori University Hospital between January 2022 and May 2022. A total of 27 patients were included in the analysis. Of these, 11 and 16 were diagnosed with mild and moderate COVID-19, respectively, based on the Japanese COVID-19 clinical practice guideline. Peripheral blood NK cell subsets and surface markers, including the activating receptor NKG2D and the inhibitory receptor TIGIT, as well as serum levels of 24 immunoregulatory markers, such as cytokines and cytotoxic mediators, were measured at admission and recovery. In addition, to explore immune patterns associated with disease severity, differences in 24 serum markers and soluble UL16-binding protein 2 (sULBP2) at the clinically most symptomatic time point during hospitalization were visualized using a volcano plot and analyzed with Spearman's rank correlation analysis and principal component analysis (PCA).Results: Patients with mild COVID-19 exhibited expanded subsets of unconventional CD56ᵈⁱᵐCD16⁻ NK cells with elevated NKG2D expression and lower levels of cytotoxic mediators (granzyme A, granzyme B, and granulysin). In contrast, patients with moderate disease exhibited NK cell exhaustion, characterized by upregulation of TIGIT, along with increased levels of NK cellassociated cytokines and cytotoxic mediators. The volcano plot identified that the patients with moderate COVID-19 exhibited significantly elevated IL-6 and sULBP2 levels. Spearman's rank correlation analysis revealed that IL-6, IFN-γ, soluble Fas, and CXCL8 were correlated with increased sULBP2. The PCA identified distinct clusters based on disease severity.The results of study highlight the differences in NK cell-associated immune alterations between mild and moderate COVID-19 cases. Elevated IL-6 and sULBP2 levels, along with their correlations with inflammatory mediators, reflects differences in immune response based on disease severity. These findings provide insight into the immune response to infection caused by the Omicron variant of SARS-CoV-2 and improve our understanding of its immunological features.