Article Figures & Data
Figures
- FIGURE 1.
Survival and changes in major blood cell populations.
(A) Survival analysis comparing overall survival of splenectomized patients with the comparator surgery patients. The difference is not statistically significant (p = 0.1846). The absolute number of platelets (B), RBCs (C), and grouped WBCs (D) per time point. The blue line indicates the splenectomized patients, and the gray line indicates the control surgery patients. The asterisks (*) indicate statistical significance between the two groups. The Roman numerals indicate statistical significance between a certain time point and baseline. Dotted lines denote normal reference range. I indicates p ≤ 0.05, II indicates p ≤ 0.01, III indicates p ≤ 0.001, and IV indicates p ≤ 0.0001. ***p ≤ 0.001, ****p ≤ 0.0001.
- FIGURE 2.
Changes in individual WBC populations: neutrophils, lymphocytes, and monocytes.
The percentage (A) and absolute number (B) of neutrophils, the percentage (C) and absolute number (D) of lymphocytes, and the percentage (E) and absolute number (F) of monocytes per time point. The percentage of the major cellular subsets per patient at time point 2–5 y postoperation (G). The blue line indicates the splenectomized patients, and the gray line indicates the control surgery patients. The asterisks (*) indicate statistical significance between the two groups. The Roman numerals indicate statistical significance between a certain time point and baseline. Dotted lines denote normal reference range. I indicates p ≤ 0.05, II indicates p ≤ 0.01, III indicates p ≤ 0.001, and IV indicates p ≤ 0.0001. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.
- FIGURE 3.
Splenectomy alters the expression of CCR4 and CCR6 on T cells and myeloid cells.
Mass cytometric analysis of lysed whole blood from patients before and after splenectomy. (C) The timing of the postsplenectomy blood draws from each patient. Files were normalized and gated on single CD45+ cells prior to analysis. (A) PhenoGraph analysis of CD45+ events comparing pre- and postsplenectomy samples from all five patients, colored by cellular phenotype (cell populations were identified based on the markers in Supplemental Table III). (B) The frequencies of PhenoGraph-defined clusters comparing the values of each cell type between pre- and postsplenectomy samples from each patient. (C) Comparison of the frequencies of PhenoGraph-defined clusters between pre- and postsplenectomy samples from each patient, including the number of days postsplenectomy the blood draw occurred. (D) The CD45+ events from pre- and postsplenectomy colored based on the expression of either CCR6 (top) or CCR4 (bottom). (E) Quantification of the mean metal intensity (relative expression level) of CCR6 (top) or CCR4 (bottom) compared between the designated cellular phenotypes and pre- and postsplenectomy samples compared across the designated cellular phenotypes.
- FIGURE 4.
Splenectomy increases the frequency of naive CD4+ T cells, activates memory or effector memory CD4+ T cells, and increases the expression of CCR6 and CCR4 on these cells.
Mass cytometric analysis of lysed whole blood from patients before and after splenectomy. (C) The timing of the postsplenectomy blood draws from each patient. Files were normalized and gated on single CD45+CD3+CD19− cells prior to analysis. (A) PhenoGraph analysis of CD45+CD3+CD19− cells comparing pre- and postsplenectomy samples from all five patients, colored by cellular phenotype. (B) The frequencies of PhenoGraph-defined clusters comparing the values of each cell type between pre- and postsplenectomy samples from each patient (cell populations were identified based on the markers in Supplemental Table III). (C) Comparison of the frequencies of PhenoGraph-defined clusters between pre- and postsplenectomy samples from each patient, including the number of days postsplenectomy the blood draw occurred. (D and E) Identification of the two populations with the greatest degree of change between pre- and postsplenectomy. (F) Dendrogram heatmap generated in Cytofkit depicting the differences in the expression of the designated proteins between clusters 21 and 22. (G) Quantification of the mean metal intensity (relative expression level) of the designated proteins compared between clusters 21 and 22 from pre- and postsplenectomy samples. *p < 0.05.
- FIGURE 5.
Splenectomy alters the frequency and activation of monocytes and neutrophils.
Mass cytometric analysis of lysed whole blood from patients before and after splenectomy. (C) The timing of the postsplenectomy blood draws from each patient. Files were normalized and gated on single CD45+CD3−CD19− cells prior to analysis. (A) PhenoGraph analysis focusing on CD45+CD3−CD19− myeloid cells, comparing pre- and postsplenectomy samples from all five patients, colored by cellular phenotype. (B) The frequencies of PhenoGraph-defined clusters comparing the values of each cell type between pre- and postsplenectomy samples from each patient. (C) Comparison of the frequencies of PhenoGraph-defined clusters between pre- and postsplenectomy samples from each patient, including the number of days postsplenectomy the blood draw occurred. (D and E) Identification of the populations with the greatest degree of change between pre- and postsplenectomy (cell populations were identified based on the markers in Supplemental Table III). (F) Dendrogram heatmap generated in Cytofkit depicting the differences in the expression of the designated proteins between clusters 2 and 14 or 15 and 24. (G) Quantification of the mean metal intensity (relative expression level) of the designated proteins compared between clusters 2 and 14 and between 15 and 24 from pre- and postsplenectomy samples. *p < 0.05.
Tables
Splenectomy (n = 95) Nonsplenectomy (n = 91) Age at surgery, y, median (range) 58 (25-85) 66 (19-88) Sex, percent female (n) 69% (66) 47% (43) BMI, median (range) 25.2 (17.4-51.6) 25.5 (16.7-38.0) Follow up, y (range) 2.67 (0.088-7.6) 4.68 (0.16-7.10) Deceased, percent (n) 26% (25) 45% (41) Cancer, n Adenocarcinoma: heada 0 47 Adenocarcinoma: tail 22 0 Neuroendocrine tumor: head 1 12 Neuroendocrine tumor: tail 20 0 Other tumorsb 17 2 Noncancer, n Pancreatic cysts 16 5 Intraductal papillary mucinous neoplasm 14 19 Other pathologiesc 5 6 Node positive, percent (n) 34% (32) 36% (34) Tumor, percent (n) Locally advancedd 19% (18) 37% (34) Distant metastasise 20% (19) 10% (9) ↵a Including adenocarcinomas of the extrahepatic bile duct, common bile duct, intrapancreatic bile duct, papilla of Vater, and duodenum.
↵b Including melanoma, ovarian cancer, endometrium cancer, colon cancer, gastrointestinal stromal tumor, and solid pseudopapillary neoplasm.
↵c Including immune thrombocytopenia, pancreatitis, pancreatolithiasis, benign vascular lesions, primary sclerosing cholangitis, and necrosis.
↵d At the time of surgery.
↵e Postsurgery.
Data Supplement
