methylcellulose and Leukocytosis

Investigation of peripheral blood cell count alterations in cases with hypersplenism, and an understanding of the relationship between splenic function and hematopoietic cell production require suitable experimental animal models. Previously described methods are either traumatic or require surgical intervention. We suggest a relatively simple method for achievement of a state mimicking hypersplenism in mice by intraperitoneal inoculation of syngeneic spleen cells. Mice were inoculated intraperitoneally with 3 x 10(7) splenocytes suspended in 0.3 ml phosphate buffered saline (PBS). After 2 months, the inoculated animals showed a progressive decrease in the peripheral white blood cell (WBC) counts and hyperplastic bone marrow that persisted until the experimental end point (7 months). Five days after inoculation of splenocytes stained with carboxy-fluorescein diacetate succinimidyl ester (CFSE), the majority of the stained cells was present in the peritoneal cavity (33%) and in the liver (13%), whereas the percentage of stained cells in the peripheral blood and the spleen cell suspension was negligible. The mitogen response of the peripheral blood mononuclear cells (PBMC) from treated mice to concanavalin A (Con A) remained unaltered. Splenocyte-inoculated mice that were further splenectomized did not show leukocytosis after splenectomy, as was observed in animals in which the spleen was removed without any pretreatment. The lack of any signs of discomfort in animals from the study group, in comparison with the visibly ill appearance and even death of mice in which hypersplenism was achieved by repeated injections of methylcellulose (MC), which served as controls, favors the convenience of the method.

Topics: Adoptive Transfer; Animals; Bone Marrow; Concanavalin A; Fluorescent Dyes; Hypersplenism; Injections, Intraperitoneal; Leukocytes; Leukocytes, Mononuclear; Leukocytosis; Liver; Methylcellulose; Mice; Mice, Inbred BALB C; Peritoneal Cavity; Spleen; Splenectomy; Transplantation, Isogeneic

22 male Sprague-Dawley rats were divided into two groups, of which group I received methylcellulose and group II saline intraperitoneally for 12 weeks. After ligation of the splenic artery of the animals in group I and sham operation of the rats in group II, the injections were continued for a further 9 weeks. At the operation, the group I animals all showed signs of hypersplenism with anemia, and leuko- and thrombocytopenia. The platelet counts normalized after the operation, a marked leukocytosis developed and the anemia was further aggravated. At the end of the study, the animals were challenged with 4 x 10(6) colony-forming units of pneumococci type 1, resulting in deaths of 11 of 12 animals in group I, in contrast to survival of all 10 rats in group II (p = 0.000017).

Topics: Animals; Hypersplenism; Leukocytosis; Ligation; Male; Methylcellulose; Pancytopenia; Pneumococcal Infections; Rats; Rats, Inbred Strains; Splenic Artery