Leukocyte migration in synovial tissue. Leukocyte distribution, orientation, and migratory pattern after immune complex deposition in rabbit knee joints.

Complement-activating bovine serum albumin (BSA)-anti-BSA immune complexes (ICs) were injected into rabbit knee joint cavities; the contralateral control joint was injected with BSA together with normal rabbit serum. The migration of leukocytes from the synovial venules into the joint cavity was analyzed with light microscopy (LM), scanning (SEM) and transmission (TEM) electron microscopy. EM autoradiography was used to study the endocytosis of ICs by leukocytes. The shape, orientation, and distribution of migrating polymorphonuclear granulocytes (PMNGs) were analyzed by LM morphometry. PMNGs accumulated in the joints injected with ICs. The peak of the number of PMNGs in the synovial tissue was reached after 4 hours, in the joint cavity after 6 hours. PMNGs in the synovial tissue were concentrated in the intimal layer. Migrating PMNGs were polarized, as judged by the ratio between the long (D max) and short (D min) axes of the cells. There was a close association between the migrating PMNGs and the collagen fibers. The morphometric data showed that the nonflattened, cylindrically-shaped PMNGs were oriented along the collagen bundles, running parallel to the synovial surface, and did not migrate in the straight direction of a theoretic leukotactic gradient originating in the joint cavity after IC deposition. SEM and TEM showed that the PMNGs were aligned along the collagen fibers and interacted activity with the collagen by pseudopods and cytoplasmic projections. EM autoradiography showed that the PMNGs in the joint cavity had ingested 125I-labeled ICs and were degranulated. In contrast, the PMNGs within the synovial membrane did not show any signs of IC endocytosis or any apparent degranulation. Synovial type A cells were found to contain ICs. This study indicates that the response of PMNGs in IC-induced synovitis consists of two distinct phases: an initial, mainly migratory phase in the synovial membrane where the PMNGs appear to use the collagen fibres as a climbing framework, and a second phase, in the joint cavity, characterized by PMNG metabolic activation, endocytosis of ICs, and degranulation. The apparent inability of PMNGs in the synovial membrane to ingest ICs and become degranulated might be due to not only concentration differences of ICs and leukotactic factors between the joint cavity and the synovial tissue but also might be related to the apparently active interaction with collagen.     

On the development of the rabbit vagina

Summary The author studied 71 female rabbit fetuses and 6 female young varying in age from 19 days p.c. to 9 days p. p. All the specimens were routinely stained in haematoxylin-eosin-orange-G after fixing in Bouin's fluid. In addition, a study was made of the occurence of beta-glucuronidase, alkaline phosphatase, and acid phosphatase in the müllerian, the wolffian, and the v.c.r. epithelium in the regions under discussion. The gradual transition between wolffian and what is here called intermediate epithelium, and the distribution of mitoses and degeneration granules, as well as the distribution of the different enzymes, suggests that the caudal part of the vagina of rabbit is derived from wolffian epithelium. The cranial part of the vagina is lined by epithelium derived from the müllerian ducts.     

Phagocytosis of spermatozoa by the rabbit vagina

Twenty-four hours after mating in the rabbit, numerous spermatozoa are observed attached to the surgace of the vaginal epithelium. Spermatozoa appear to be attached by their heads to microvilli of the simple columnar cells which compose the vaginal epithelium. Spermatozoa are taken up by the epithelial cells, and they are found within the cells in various stages of degeneration up to seven days after mating. Epithelial cells frequently appear to be filled with numerous vacuoles containing sperm components in various stages of degeneration. It is suggested that some property of the surface of sperm heads may render them particularly susceptible to phagocytosis. Phagocytosis of spermatozoa may not have long-term harmful effects for the epithelium since these epithelial cells presumably have a high rate of turnover.     

Dextran sulphate inhibits neutrophil emigration and neutrophil-dependent plasma leakage in rabbit skin

Rolling of neutrophils in postcapillary venules is believed to be an obligatory step in the adhesion of neutrophils to endothelial cells, preceding firm attachment and emigration. This rolling on the endothelial surface can be blocked by sulphated polysaccharides. Therefore, dextran sulphate or dextran T500 was injected intravenously (i.v., 25 mg/kg) and neutrophil infiltration and plasma leakage in skin were measured in response to intradermal (i.d.) injection of inflammatory mediators in the presence of calcitonin gene-related peptide (CGRP). Neutrophil accumulation and oedema formation induced by the neutrophil chemoattractants, f-Met-Leu-Phe (FMLP) or C5adesArg, were completely suppressed in the presence of dextran sulphate. Neutrophil emigration in response to interleukin-1 (IL-1) was not affected by dextran sulphate. These results suggest that neutrophil recruitment induced by FMLP and C5adesArg, but not by IL-1, is mediated via sulphated glycans, perhaps on endothelial cells.     

Leukocyte migration inhibition test in patients with trophoblastic tumors

Inhibition of leukocyte migration under the influence of protein extract from a chorionepithelioma was investigated in 40 patients with trophoblastic tumors. Marked inhibition of leukocyte migration was found in patients with evidence of active tumor growth before and in the course of treatment. The exception was four patients in whom no inhibition was found either before or during treatment. The inhibition effect was absent in mos patients when the clinical course was favorable as a result of surgery and chemotherapy or of chemotherapy alone. Leukocytes of healthy donors did not react to the tumor extract in any of 24 cases.Group for Combined Methods of Treatment, Department of Gynecology, Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 5, pp. 580–582, May, 1978.     

Leukocyte migration test in a modified agarose medium.

The medium used for the leukocyte migration test contained 1% agarose, 0.003 M HEPES, TC medium 199, and 10% horse serum. The pH was adjusted to 7.3 with 2 N NaOH. The leukocyte migration test in this medium correlated with the intradermal test in 36 subjects when PPD was used as antigen, and the results were comparable to those of the reference technic. Addition of HEPES to the original medium enables the test to be performed in a standard incubator.     

Leukocytes in chemotactic-fragment-induced lung inflammation. Vascular emigration and alveolar surface migration.

Lung inflammation was induced in rabbits by intratracheal injections of chemotactic fragments obtained from zymosan-activated serum (CF-ZAS), and the route of vascular emigration and alveolar surface interaction of polymorphonuclear leukocytes (PMNs) and monocytes migrating into the lung was characterized by transmission (TEM) and scanning (SEM) electron-microscopic examination. Leukocytes migrated from capillaries and venules into the alveolar wall interstitium by adherence to the vascular endothelium and migration through the endothelial intracellular junction to attain a position between a reapposed endothelial cell junction and the vascular basement membrane. The cells then migrated into the interstitium through a narrow opening in the basement membrane. Leukocyte entrance into the alveolar space from the interstitium appeared to occur through small openings in the epithelial basement membrane at or near the Type I epithelial intercellular junction. Once in the alveolus, PMNs and macrophages demonstrated surface adherence and spreading along with evidence of migration, pseudopod extension, interalveolar pore transit, and retraction fiber formation. This study indicates the leukocyte influx into the alveolus in acute chemotactic-factor-induced inflammation is via a continuum of migrational activity, beginning at the pulmonary capillary endothelial surface and persisting on the alveolar epithelial surface.     

Leukocyte migration inhibiting factor in preparations of alpha and gamma interferons

High titres of leukocyte migration inhibition factor (LMIF) activity were found in native and partially purified preparations of human alpha- and gamma-interferon. In the course of chemical purification of interferon up to 98% of LMIF was lost. The highest content of LMIF was in gamma-interferon preparations, while recombinant alpha 2-interferon (reaferon) showed no LMIF activity.     

Leukocyte migration inhibition test in children with cow milk allergy

The usefulness of the leukocyte migration inhibition factor (LIF) test to detect cow milk (CM) hypersensitivity was studied in 40 children with suspected allergy to CM. Hypersensitivity was carefully investigated by oral milk challenges, which gave a final confirmation of cow milk hypersensitivity in 12 subjects, and excluded it in the remaining 28 subjects. Leukocyte migration inhibition was measured using beta-lactoglobulin (BLG), alpha-lactalbumin (ALA), alpha-casein (ACA) and bovine serum albumin (BSA) as antigens. IgA and IgG antibodies to these antigens were measured by an enzyme-linked immunosorbent assay, and IgE antibodies to these antigens and to CM by radioallergosorbent test (RAST). Skin prick test with CM was performed in 38 subjects, and with BLG, ALA, ACA and BSA in 29 subjects. Leukocyte migration was more often inhibited by cow milk antigens in the CM challenge positive (CM+) subjects than in the challenge negative (CM-) subjects. Of the specific milk antigens, ALA was the most potent inhibitor, and gave a positive LIF test result in all CM+ subjects, and significantly (P less than 0.02) less often (15/24) in CM- subjects. Also in the skin prick test and RAST, ALA gave positive results more often than the other milk antigens. BLG, ACA and BSA had an inhibiting effect on leukocyte migration, but the difference between the CM+ and CM- subjects was not statistically significant. Two of the 12 CM+ subjects had no demonstrable IgE antibodies to CM proteins; both of them, however, had a positive LIF test with at least one of the CM antigens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Viability and freezing ability of rabbit embryos collected in the vagina after prostaglandin treatment

Morulae were collected from rabbit vaginas after prostaglandin treatment 65h post coitum. The optimum embryo recovery was obtained when the flushings started around 12h after the prostaglandin injection. The mean embryo collection was around 10 embryos per animal. These embryos had the same viability as those collected by the classical technique of uterine flushing (around 80%). The freezing ability of these embryos was also similar to that observed for uterus-collected embryos. Embryos not ejected after the treatment can develop in vivo and give birth to living progeny. The results obtained suggest that this technique can be used for egg transfer in rabbit genetic improvement programs.     

Post-operative consequences of glove powder used pre-operatively in the vagina in the rabbit model

This study was undertaken to determine if a retrograde migration of glove powder (Biosorb(TM)) from the vagina into the abdominal cavity in combination with a subsequent intra-abdominal trauma could affect the development of adhesions. A rabbit model was used with one control and one test group. A total of 50 mg Biosorb(TM) was deposited intravaginally in the test cases before ovulation was induced. Three days later a laparotomy with a small standardized surgical trauma was carried out on the left Fallopian tube and the ipsilateral peritoneum. Ten days later the extent of intra-abdominal adhesions was evaluated. The presence of starch particles in cell smears and biopsies was also investigated. Significant differences in the formation of adhesions were found between the control and the test cases (P < 0.001). In the test group there were dense adhesions and in four cases the Fallopian tube was completely attached to the peritoneal wall. In the control cases only loose minor adhesions were found. This study indicates that the number of starch particles migrating from the vagina to the abdominal cavity is sufficient to enhance significantly the formation of post-operative adhesions. We therefore suggest that powder-free medical or surgical gloves should be used in obstetrics and gynaecology.     

In-vivo effects of anti-inflammatory and other drugs on granulocyte emigration in the rabbit skin collection chamber.

A method for measuring localised leukocyte mobilisation under in-vivo conditions with a plastic skin collection-chamber adapted to the rabbit ear was used for assessing the effects of anti-inflammatory and other agents on granulocyte emigration. Studies on the effect of oral drug administration to rabbits indicated that most anti-inflammatory drugs, two cystostatic agents (cyclophosphamide and colchicine), but none of the other compounds exhibiting antihistaminc, beta-adrenolytic or neuroleptic properties, significantly inhibited granulocyte mobilisation. The results after topical application of some of these agents into the chamber correlated well with those obtained after oral treatment. This technique may thus prove useful in selecting new compounds inhibiting granulocyte mobilisation in acute inflammatory reactions.     

Leukocyte behaviour and permeability in the rat mesenteric microcirculation following induction of ovulation

BACKGROUND: Ovarian hyperstimulation syndrome (OHSS), a highly dangerous and incompletely understood complication of ovulation induction with exogenous gonadotrophins, can include haemoconcentration, hypovolaemia, hypotension, acute renal insufficiency, thromboembolism and ultimately death. Using intravital microscopy, we examined microvascular permeability and leukocyte-endothelial cell interactions in the rat mesenteric microcirculation associated with induction of ovulation. METHODS: In female rats treated with hMG and hCG, mesenteric venules were observed by intravital microscopy assisted by a video imager. Erythrocyte velocity was monitored, and rolling and adhesion of leukocytes were studied by transmission video images. Transvascular leakage of fluorescein isothiocyanate-labelled albumin was assessed by epi-illumination. RESULTS: Administration of hMG and hCG significantly increased vascular protein leakage within a few hours, and also reduced rolling velocities of leukocytes in venules and increased numbers of leukocytes adherent to endothelium at 16 h following hCG injection. The administration of antibodies against intracellular adhesion molecule (ICAM)-1 inhibited these reactions. CONCLUSION: By induction of ovulation, vascular permeability is increased not only at the surface of the ovary but also in the mesentery. Alteration of leukocyte behaviour in the microcirculation through mechanisms involving ICAM-1 is one likely cause of the protein leakage.     

Morphologic changes in the rat uterus following natural mating and embryo transfer

In order to gain some insight into the putative immune suppression that may be induced at the placental implantation sites, the morphological changes at these sites following natural matings and following the transfer of embryos fertilized in vivo were studied. The only histologic parameter that showed a significant difference was the number of granulated metrial gland (GMG) cells. More GMG cells were present in allogeneic than in syngeneic pregnancies, and more GMG cells were present following embryo transfer into an allogeneic female than following the comparable natural mating. The role of the GMG cells in pregnancy is, however, still unresolved.     

DNA replication in the epithelium of rat vagina and lower cervix following estrogen-progesterone treatment

The purpose of this study was to ascertain the effects of different amounts of progesterone on estrogen-induced DNA synthesis in the vaginal and lower cervical epithelia of the rat. Ovariectomized rats were injected subcutaneously with 1 μg of estradiol-17β dipropionate or with estradiol and 1, 5, 10 or 15 mg of progesterone for three days; control rats received oil. Tritiated thymidine was injected one hour prior to necropsy; the tissues were processed for autoradiographic study and the percentages of labeled nuclei in the basal layer from the lower half of both the vagina and cervix were determined. In the group of rats given estradiol and 1 mg of progesterone the thymidine uptake in the vagina was significantly increased over that of rats given only estrogen. However, in the rats treated with estradiol and 5, 10 or 15 mg of progesterone, the extent of DNA replication in the basal epithelial nuclei was similar to the estrogen group. These responses were compared with those of the basal epithelium from the lower half of the cervix. None of the estrogen-progesterone treatments increased the thymidine index in the cervical epithelium above that of estrogen alone; additionally, 5, 10 or 15 mg of progesterone given with estrogen suppressed DNA replication. These data indicate that stratified epithelia of these two portions of the reproductive tract respond differently to exogenous ovarian hormones.