Peritoneal Cell Preparations Research Articles

Effect ofin vivocorticosterone and acute food deprivation on rat resident peritoneal cell chemiluminescence after activation ex vivo

Adrenoglucocorticoid regulation of rat peritoneal monocyte/macrophage function was studied by exposing rats to corticosterone (CS) in the drinking water, and to fast (48 h). Production of reactive oxygen metabolites was measured by luminol amplified chemiluminescence (CL) in preparations of peritoneal cells activated by serum treated zymosan (STZ). Administration of CS which led to an increase in plasma CS from 31 (controls) to 46 ng mL-1, reduced CL (per cell) by 31%. Fast, which did not change plasma CS or ACTH, also had an inhibitory effect on CL (-25%), while the combination of CS administration and fast strongly inhibited the CL (-89%), indicating that plasma CS and fast reduced CL in a synergistic way. Similar effects on cell number were observed: CS-administration, fast and the combination reduced macrophage numbers (-13, -19.7 and -55%), while no significant effect was observed on the number of monocytes. The effect of adrenalectomy (adx) was studied in another series of experiments; adx induced no significant change in peritoneal leucocyte number or composition, while cells from adx animals had significantly higher chemiluminescence reaction than cells from sham operated animals. CS substitution in adx animals reduced CL by 30% while sham operated animals had 49% lower CL in adx. The data from adx animals also suggest that endogenous levels of CS are inhibitory for CL, but the results are not conclusive for the effect of very low doses of CS since other mechanisms than elimination of CS could prime the chemiluminescence reaction after adx. In conclusion, a moderate elevation of CS after systemic administration in vivo reduced the total number of mononuclear phagocytes in rat peritoneum, reduced the relative number of macrophages compared with monocytes, and suppressed the function of monocytes/macrophages by reducing the production of reactive oxygen molecules in activated cells. Furthermore, the effect of corticosterone was also dependent on the physiological situation, since the effects of fast and corticosterone were synergistic.

Role of Dialysis Modality in Responses of Blood Monocytes and Peritoneal Macrophages to Endotoxin Stimulation

Constitutive and endotoxin (lipopolysaccharide [LPS])-stimulated release of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and prostaglandin E2 (PGE2) by blood monocytes and peritoneal cell preparations from patients on various forms of dialysis was measured. Monocytes were obtained from healthy controls (n = 20), and from patients on peritoneal dialysis (n = 8), on hemodialysis with cellulose ester membranes (n = 9), and on hemodialysis with polysulfone membranes (n = 8). Peritoneal macrophages were recovered by lavage during laparoscopic surgery from 11 healthy controls, from dialysate in 37 patients on peritoneal dialysis, and at catheter placement for transfer to peritoneal dialysis from eight patients on hemodialysis with polysulfone membranes. Peak LPS-induced production of TNF-alpha, IL-1 beta, and IL-6 by monocytes from patients on peritoneal dialysis and cellulose ester hemodialysis was less than that by monocytes from healthy controls. In contrast, monocytes from patients treated with polysulfone hemodialysis produced amounts of cytokine not different from controls. Lipopolysaccharide-stimulated PGE2 production by monocytes was the same in both patient and control groups. Peritoneal cell preparations from patients on peritoneal dialysis showed decreased release of IL-1 beta and TNF-alpha as compared with control peritoneal cells and with their own blood monocytes. To determine whether the decreased response to LPS by peritoneal cells compared with their own blood monocytes could be attributed to lower numbers of macrophages in the peritoneal cell preparations, adherence of peritoneal cells to plastic was performed. Adherence increased the percentage of macrophages from 70% to more than 90%. In monocytes and adherence purified peritoneal macrophages from peritoneal dialysis patients, no significant difference between monocytes and adherent peritoneal macrophages could be found for TNF-alpha and PGE2. Interleukin-1 beta production by the adherent peritoneal macrophages, as by total peritoneal cells, was significantly lower than that by monocytes. Constitutive production of PGE2 and IL-6 by peritoneal cells from patients on peritoneal dialysis was significantly increased. In contrast, LPS-stimulated production of TNF-alpha, IL-1 beta, and IL-6 by blood monocytes and peritoneal cells from patients receiving hemodialysis with polysulfone membranes was comparable to that produced by monocytes and peritoneal cells obtained from healthy controls. Thus, blood monocytes and peritoneal cells from patients on peritoneal dialysis and from patients on hemodialysis with cellulose-ester membranes demonstrate a decreased cytokine response to LPS, suggesting a state similar to endotoxin tolerance.(ABSTRACT TRUNCATED AT 250 WORDS)

Binding sites for short-term glycated albumin on peritoneal cells of the rat

The interaction of in vitro short-term glycated rat serum albumin with rat peritoneal cells (40% macrophages) was investigated. Using 125I-labeled albumins the following results were obtained. Glycated albumins showed a binding reaction at 4°C, which appeared to reach equilibrium within 2 h. The concentration-dependent binding of glycated albumin showed saturation. Binding data evaluated for glycated albumin using the Sips equation are: average association constant K o = 3.15 · 10 7 M −1 with a heterogeneity index of a = 0.8 and 1.12 · 10 4 binding sites per cell. Such binding sites were identified in 40% of the peritoneal cell preparations studied. Native albumins, maleylated albumin, chondroitinsulfates, polylysine, lysine, fructose, glucose and hexitol-lysine could not compete with radio-labeled glycated rat albumin for its binding site on peritoneal cells. Effective competitors were glycated human serum albumin, glycated polylysine and fructose-lysine. Although the contamination with minute amounts of advanced glycosylation end products (AGE) could not be excluded, short-term glycated albumin was found to be bound to membranes of peritoneal phagocytotic cells by fructose-lysine specific proteins, whose approximately defined molecular masses of 290 kDa are distinct from hitherto described binding proteins for AGE- and aldehyde-modified proteins or for the scavenger receptors.

Failure of dietary restriction to enhance cytolytic and chemiluminescent activity of peritoneal exudate cells from ageing rats

Investigations into the cytolytic activity of peritoneal macrophages from male Sprague-Dawley rats (activated in vivo with Corynebacterium parvum) utilised SV40-3T3 and L-929 target cells in a 72 h [ 3H]thymidine release assay. The dietary restricted rats under test were given a measured amount of food from weaning sufficient to maintain their body weights at approximately 50% of age-matched controls fed ad libitum. Cytolytic activity was similar in the two dietary groups at both 13 and 21 months of age, nor were chemiluminescent responses of peritoneal cells to latex or zymosan particles affected by the dietary restriction. No differences were seen in the effectiveness of zymosan particles opsonised with serum pooled from dietary restricted versus control rats in stimulating chemiluminescent responses by standard preparations of peritoneal cells, and concentrations of serum C3 and IgG appeared to be unchanged by the dietary regime.

In vitro toxicity of T-2 mycotoxin in mouse lymphoid cells.

The in vitro toxicity of T-2 toxin towards mouse lymphoid cells prepared from spleen, thymus, peritoneal lavage and bone marrow cells was studied. Bone marrow cells were more resistant to damage by T-2 toxin than thymus, spleen and peritoneal cell preparations.

The measurement of histamine release from suspensions of rat peritoneal cells rich in mast cells.

Histamine released from the mast cells in unfractionated rat peritoneal cell suspensions could be quickly and conveniently measured by an automated chemical method. There were no substances in the unfractionated peritoneal cells that interfered with the chemical histamine measurement. Organic extraction of histamine and deproteinization of samples were not necessary using the automated method. The amount of histamine released from preparations of peritoneal cells by a fixed concentration of compound 48/80 decreased with the time of preincubation of the cells but this varied between preparations. Phagocytic activity directed against the mast cells probably explained these observations. The state of nutrition of the rats and the presence or absence and/or glucose in the medium all influenced the rate of decline of viability of the mast cells.

In vitro migration of peritoneal and spleen cells and its inhibition in some avion species

Cell migration and its inhibition was tested by the capillary tube technique with peritoneal exudate cells and spleen cells of chicken, turkey, goose, guinea fowl, and Japanese quail. Peritoneal cells were produced by ip administration of proteose peptone and harvested 24 hr later. Liquid paraffin proved to be unsatisfactory for preparation of peritoneal cells in some avian species. Mononuclear cells represented no more than 50–60% of the peritoneal cell populations, the other 50% being polymorphonuclear cells in all five avian species studied. Cell migration was demonstrated with chicken, turkey, and goose peritoneal and spleen cells, but not with those of guinea fowl and Japanese quail. The composition of the cell populations in the migration areas was nearly the same as in the initial preparations of peritoneal and spleen cells. Spleen cell migration was inhibited to a greater extent than that of peritoneal cells. Migration inhibitory factor (MIF) produced by chicken and turkey lymphocytes exhibited some species specificity.

Hyporeactivity due to infection: recognition of a transferable hyporeactive factor in the serum of encephalomyocarditis virus-infected mice.

As previously reported, mice infected with encephalomyocarditis (EMC) virus progressively develop a state of hyporeactivity to interferon induction as evidenced by an 80% reduction in serum interferon levels produced in response to four different inducers administered 96 h after initiation of infection. In the current report, this hyporesponsiveness was further documented in vitro in peritoneal exudate cells harvested at the time of maximal hyporeactivity (96 h) during EMC virus infection. These peritoneal cell preparations (containing no detectable interferon or virus) produced significantly less interferon than cells from uninfected mice following exposure to Newcastle disease virus or Chikungunya virus. More importantly, hyporeactivity could be transferred to normal peritoneal cells incubated in serum (containing no detectable interferon or virus) from EMC virus-infected mice during the hyporeactive period. Incubation of peritoneal exudate cells with up to a 1:200 dilution of serum from hyporeactive animals still resulted in a significant reduction in the interferon response induced by Newcastle disease virus or Chikungunya virus. Furthermore, it could be shown that this serum hyporeactive factor was effective in mouse cells but not in rabbit, chicken, or human cells.