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. 2023 Mar;44(3):296-305.
doi: 10.15537/smj.2023.44.3.20220873.

Phenotypical evaluation of lymphocytes and monocytes in patients with type 2 diabetes mellitus in Saudi Arabia

Mamdouh Allahyani  1 , Amani M Alshalawi  1 , Maram R Alshalawii  1 , Shahad A Alqorashi  1 , Abdulelah Aljuaid  1 , Mazen M Almehmadi  1 , Mohammed A Bokhary  1 , Alhanouf S Albrgey  1 , Ahmad A Alghamdi  1 , Abdullah F Aldairi  1 , Ayman S Alhazmi  1
Affiliations

Phenotypical evaluation of lymphocytes and monocytes in patients with type 2 diabetes mellitus in Saudi Arabia

Mamdouh Allahyani et al. Saudi Med J. 2023 Mar.

Abstract

Objectives: To evaluate the levels of total lymphocytes, B-lymphocytes (CD19+), T-lymphocytes (CD3+), natural killer (NK) cells (CD3-/CD56+), and monocyte subsets in type 2 diabetes mellitus (T2DM) patients in Saudi Arabia. In addition, this study aimed to evaluate whether B- and T-lymphocyte subsets are frequently altered in patients with T2DM.

Methods: A case-control study included 95 participants recruited in the study: 62 patients with T2DM and 33 healthy individuals. All the patients were admitted to the Diabetic Centre in Taif, Saudi Arabia. Blood samples were collected between April and August 2022. The hemoglobin A1c (HbA1c) level was evaluated in all patients. Flow cytometry was used to measure the expression of B-lymphocyte, T-lymphocyte, NK cells, and monocyte markers. The unpaired t-test was carried out to evaluate the differences in these markers between T2DM patients and healthy individuals.

Results: Patients with T2DM were associated with a lower percentage of total lymphocytes, higher percentage of B-lymphocytes, naive, and memory B subsets. In addition, patients with T2DM showed lower percentage of total T-lymphocytes (CD3+) and CD4 T-cells, but higher CD8 T-cell expression. Also, the NK-cell level was reduced in patients with T2DM, and the levels of monocyte subsets were altered.

Conclusion: These data suggested that levels of lymphocytes and monocytes are impaired in T2DM patients, and this might be associated with the higher risk of infections observed in these patients.

Keywords: T2DM; lymphocytes; monocytes.

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Figures

Figure 1
Figure 1
- Percentage of lymphocytes in T2DM patients and healthy individuals. Lymphocytes were isolated from the peripheral blood of the T2DM patients and healthy individuals based on SSC and FSC using flow cytometry. A) Dot plots showing the percentage of lymphocytes in the samples obtained from healthy individuals and T2DM patients. B) The percentage of lymphocytes in both groups is presented. Data were analyzed using an unpaired t-test. Values are presented as mean±SD. *** p≤0.001, T2DM: type 2 diabetes mellitus, SSC: side scatter, FSC: forward scatter
Figure 2
Figure 2
- Levels of B-lymphocytes and their subsets in T2DM patients and healthy individuals. Lymphocytes were isolated from the peripheral blood of the T2DM patients and healthy individuals. The cells were then stained with anti-CD19 (FITC), CD27 (PerCyp Cy5.5), and CD38 (APC) and analyzed using flow cytometry. A) Representative dot plots showing the percentage of CD19+ B-cells in the samples obtained from healthy individuals and T2DM patients. B) The percentage of CD19+ cells in both groups is presented. C) Representative dot plots showing the percentage of B-cell subsets in the samples obtained from healthy individuals and T2DM according to CD27 and CD38 expression. D) The percentage of CD27-/CD38+ cells in both groups is presented. E) The percentage of CD27+/CD38- cells in both groups is presented. Data were analyzed using an unpaired t-test. Values are presented as mean±SD. *** p≤0.001, **** p≤0.0001, T2DM: type 2 diabetes mellitus
Figure 3
Figure 3
- Levels of T-lymphocytes and subsets in T2DM patients and healthy individuals. Lymphocytes were isolated from the peripheral blood of T2DM patients and healthy individuals. The cells were then stained with anti-CD3 (APC Cy7), CD4 (APC), and CD8 (PE) and analyzed using flow cytometry. A) Dot plots showing the percentage of CD3+ T-cells in the samples obtained from healthy individuals and T2DM patients. B) The percentage of CD3+ cells in both groups is presented. C) Dot plots showing the percentage of CD4 and CD8 T-cells in the samples obtained from healthy individuals and T2DM patients. D) The percentage of CD4+ cells; and E) CD8+ cells in both groups is presented. Data were analyzed using an unpaired t-test. Values are presented as mean±SD. * p≤0.05, T2DM: type 2 diabetes mellitus
Figure 4
Figure 4
- Levels of NK cells in T2DM patients and healthy individuals. Lymphocytes were isolated from the peripheral blood of the T2DM patients and healthy individuals. The cells were then stained with anti-CD3 (APC Cy7), and CD56 (PE) and analyzed using flow cytometry. A) Dot plots showing the percentage of CD3-/CD56+ NK cells in the samples obtained from healthy individuals and T2DM patients. B) The percentage of NK cells in both groups is presented. Data were analyzed using an unpaired t-test. Values are presented as mean±SD. ** p≤0.01, T2DM: type 2 diabetes mellitus
Figure 5
Figure 5
- Expression of monocyte subsets in T2DM patients and healthy individuals. Monocytes were isolated from the peripheral blood of the T2DM patients and healthy individuals. The cells were then stained with anti-CD14 (APC Cy7), and CD16 (FITC) and analyzed using flow cytometry. A) Dot plots showing the percentage of monocyte subsets in the samples obtained from healthy individuals and T2DM patients. B) The percentage of CD14+ cells; C) CD14+/CD16+; and D) CD16+ in both groups are presented. Data were analyzed using an unpaired t-test. Values are presented as mean±SD. * p≤0.05, *** p≤0.001. T2DM: type 2 diabetes mellitus
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