Age-related production of spotted souslik (Spermophilus suslicus) interferon

Interferon production by virus stimulated spotted souslik tissue explants in vitro depends on the age of the tissue donor and on the kind of tissue. The interferon titers in foetal liver were 1/120-1/370 and foetal lungs 1/21-1/500 of these observed in autologous organs of adult sousliks after infection in vitro with Newcastle disease virus, Radom strain (NDV-R). After birth interferon response in vivo develops with the age. Nine-day-old suckling sousliks were not able to respond with interferon production to lipopolysaccharide (LPS), but serum interferon levels after intraperitoneal (i.p.) injection of polyinosinic polycytidylic acid complex (poly I. poly C) complexed with diethyl-amino-ethyl-dextran (DEAE-dextran) or after injection of NDV-R were nearly similar to those in adult animals. In tissue however, levels of interferon were always lower than in adults. Young 4-5-week-old sousliks responded to induction with NDV-R and poly I. poly C with DEAE-dextran and tilorone hydrochloride as adults and produced interferon in serum and different organs, however, after induction with LPS serum interferon level did not reach the values observed in serum of adult sousliks.     

Fetal and adult bovine interferon production during bovine viral diarrhea virus infection.

Levels of interferon in adult bovine serum and in fetal bovine serum and tissues were examined during the course of transplacental bovine viral diarrhea virus infection. The cows produced circulating interferon between 2 and 9 days after viral inoculation, with mean peak levels in the serum on day 4. Interferon could be routinely detected in fetal tissues (e.g., thymus, spleen, and kidney) between days 4 and 21 after viral inoculation of the cows at 149 to 150 days of gestation (mid-second trimester) and in fetal serum from day 13 through day 21. Interferon was also detectable in the serum and tissues of fetuses from dams infected at day 95 of gestation (the beginning of the second trimester). In general, no differences were found between the ability of the adult and fetus to produce interferon. Fetal lamb kidney cells were more sensitive to the antiviral effects of bovine interferon than were fetal bovine kidney cells. The antiviral substance from the fetal and adult animals was characterized as interferon by standard criteria.     

Effect of interferon and poly I:C on replication of influenza virus in organ cultures of human origin.

Interferon and poly I : C inhibited the growth of influenza virus in organ cultures of foetal nasal mucosa, trachea, oesophagus and adult nasal mucosa. No antiviral effect was observed in decidua organ cultures. The combined use of interferon and poly I : C resulted in the enhancement of the antiviral effect.     

Production of lymphokines and interferon by immune cells involved in recovery of mice from herpes simplex virus type 2 hepatitis

Adoptive transfer of spleen cells from mice 4 days after infection with herpes simplex virus type 2 (HSV-2) reduced the virus titer in the liver of recipient mice infected 24 h before transfer. Macrophage chemotactic factor (CF) and macrophage migration inhibition factor (MIF) were produced by day 3 of infection in spleen cell cultures stimulated with HSV-2, but not with control antigen, i.e. 1 day before the cells are active in adoptive transfer. Interferon was produced in cultures established throughout the infection but not in normal spleen cells. From days 1 to 5 of infection interferon was produced irrespective of in vitro restimulation, although the highest amounts were always produced after stimulation with the specific antigen. Spleen cells from mice infected for 6 days produced interferon only when stimulated with HSV-2. The cells from 6-day-immune mice active in adoptive transfer and CF and MIF production were found to be Thy 1+, Ig- and Lyt2-. Both Thy 1+ and plastic adherent cells were necessary for interferon production, whereas Ig+ and Lyt2+ cells did not produce interferon. The interferon was acid stable and neutralized by antiserum against alpha/beta-interferon and thus has the characteristics of alpha-interferon. The data indicate that a delayed type hypersensitivity reaction with lymphokine-induced macrophage recruitment into infectious foci may be a central feature of the recovery process in HSV-2-induced hepatitis. A possible role of interferon produced by the accumulated cells needs further investigation.     

In vitro production and cellular origin of murine type II interferon.

Antigen-specific type II interferon was produced in vitro by harvesting supernatants of spleen cell cultures from Swiss-Webster mice sensitized with Mycobacterium bovis strain BCG and challenged with old tuberculin. Treatment of C3H mouse spleen cell cultures with appropriate anti-Ia, anti-IgG, anti-Thy-1 or anti-Ly-2,3 sera resulted in a significant decrease in production of type II interferon. Removal of nylon wool adherent cells or cells with histamine receptors by column chromatography similarly caused reduced production of type II interferon. Recombination of spleen cell cultures treated with anti-Ia and anti-Thy-1 sera or of cells treated with anti-IgG and anti-Thy-1 resulted in restored production of type II interferon. Interferon production was also restored by combination of cells passed through histamine columns with anti-Ia treated cells, or those passed through nylon wool columns with anti-Thy-1 treated cells. Anti-Ly-1 serum treatment had no effect on interferon production. Removal of plastic-adherent cells or cells that had phagocytosed carbonyl iron also decreased interferon production, suggesting that macrophages were also involved in type II interferon production. Recombination of non-adherent spleen cells with anti-Ia and anti-Thy-1 sera treated spleen cells, however, did not restore interferon production, suggesting that other cells in addition to macrophages are depleted by the adherence procedure. These findings indicate that type II interferon is produced by suppressor or cytotoxic (Ly-2,3+) T lymphocytes in co-operation with one or two additional cell types: (i) B lymphocytes, and (ii) macrophages.     

The ability to predict weight gain,individual organ weight,and corresponding food intake in the rat by the four-parameter model for physiological responses

The applicability of the four-parameter model for physiological responses to the prediction of food intake and corresponding weight gain and individual organ weight gain was studied further in 40-day postpartum male rats. Seven groups of animals were maintained on diets in which protein content ranged from 0 to 23.54% casein. Food intake and weight gain were recorded every other day for each animal for 21 days. At the termination of the experiment the following organs were removed and weighed: liver, heart, lungs, spleen, kidneys, adrenals, and testes. When these weight values are fitted by use of the four-parameter model, food intake and total animal and organ weight gains can be predicted in relation to the amount of protein in the diet. It was found that liver, heart, lungs, spleen, and whole animal had similar K(0.5) values. However, it was also shown that there is variation in response of organs when relating organ weight as a percentage of body weight. For example, heart, lungs, and testes show an increased ratio on low protein diet while liver, kidneys, and adrenals maintain a fairly constant ratio and the spleen shows a decreased ratio. Additionally, it was noted that the animals on low protein diet consumed more food per gram body weight but did so at a slower rate. Possible future applications of the four-parameter model for physiological reponses are discussed.     

A comparative study of the fixation of antibiotics of the tetracycline group by homogenates of organs

Summary The authors established the quantitative fixation of chlortetracycline, tetracycline, and oxytetracycline by homogenates of the liver, kidneys, spleen, lungs, heart, and brain of white rats. Fixation by the organ homogenates was greatest by chlortetracycline, followed by tetracycline and oxytetracycline in that order. Homogenates of the liver, kidneys, and brain fixed the tetracyclines in considerable quantitities, those of the lungs, heart and spleen—to a noticeably lesser degree.Presneted by Active Member AMN SSSR V. V. Zakusov     

Interferon Production by Corynebacterium Parvum and BCG-Activated Murine Spleen Macrophages

Macrophages were identified to be a major source of interferon produced in murine spleen cell cultures after intravenous injection of Corynebacterium parvum (C. parvum), strain CN 6134 or Bacille Calmette Guérin (BCG). Another strain of C. parvum, CN 5888, which lacks RES stimulating activity and adjuvant activity in vivo, was not effective when injected intravenously. Protein synthesis was required for interferon activity to be produced and protein synthesis was also required for the antiviral state to be expressed. The antiviral activity was relatively stable to pH 2 and neutralized by an antiserum against virus-induced fibroblast interferon, thus exhibiting some properties of type I interferon. In vitro only CN 6134, the biologically active strain, could induce small amounts of interferon in spleen macrophage cultures. Macrophages from CN 6134 or BCG-infected athymic nu/nu mice produced similar interferon titers as their controls. It is concluded that infection with certain immunomodulators can activate splenic macrophages via a predominantly T-cell independent mechanism. Interferon in turn may operate locally as a mediator of immunoregulation.     

Influence of divinyl ether-maleic anhydride (pyran) on foot-and-mouth disease virus infection: Effect on adsorption and multiplication in mouse tissues

Summary The mechanism of resistance to foot-and-mouth disease virus (FMDV) which occurs 2 days after divinyl ether-maleic only anhydride (pyran copolymer) treatment but diminishes by the 7th day was investigated. Adsorption of virus by minced infant mouse spleen, muscle, kidney, heart or liver was not affected either at 2 days or 7 days after treatment of the mice with pyran. However,in vitro multiplication of FMDV in minced kidney was affected by pyran at both times. The resistance of mice 2 days after treatment was related to absence of virus in muscle, kidney, and spleen tissues. Seven days after pyran administration, however, FMDV was recovered from muscle and spleen but not kidney. Interferon was recovered from various tissues 2 days but not 7 days after mice were injected with pyran. Other unknown polymer-induced antiviral factors as well as interferon may have been related to the resistance mechanism.     

Development of B-lymphocyte colony-forming cells in foetal mouse tissues.

In CBA and (CBA X C57B1)F1 mice, cells forming B-lymphocyte colonies in agar culture were first detected in the 17-day foetal liver and the following day in the spleen, bone marrow and peripheral blood. Colony-forming cells were not detected in the yolk sac or foetal thymus. Adult levels of colony-forming cells were achieved within 3 days of birth. In organ cultures of 15-day foetal liver or spleen, B-lymphocyte colony-forming cells developed during a 5-day incubation period, indicating that both organs can function as bursal analogues. Foetal liver colony-forming cells were of small size and generated colonies of cells with a pattern of membrane immunoglobulin similar to colony cells generated by cells from adult animals.     

Histopathologic and ultrastructural studies of disseminated cytomegalovirus infection in strain 2 guinea pigs

Inbred strain 2 guinea pigs developed severe disseminated disease during acute experimental guinea pig cytomegalovirus (GPCMV) infection. A high mortality rate (100%) resulted, with most animals dying between 10 and 14 days after high dose (7.5 X 10(5) TCID50) virus inoculation. Infectious virus was recovered from many tissues, including spleen, lungs, liver, pancreas, heart, adrenals, kidneys, and salivary glands. The rate of GPCMV isolation from these tissues ranged from 50 to 100%. Gross lesions were observed in the spleen, liver, and lungs. On histologic examination, lesions were also seen in many other organs, including heart, pancreas, kidneys, adrenals, brain, intestines, and salivary glands. Intranuclear viral inclusions were present in many cell types of various organs. Under electron microscopic examination, cells with viral inclusions were easily found in the spleen, and liver, but less readily in the lungs, kidneys, salivary glands, and other organs. Most of the intranuclear inclusions consisted of electron-dense fibrils (10 nm diameter), viral nucleocapsids (100 nm), and tubular structures (60 nm diameter). Dense bodies and enveloped dense virions containing single or multiple capsids were present in the cytoplasm of many infected cells. The morphologic developments of GPCMV in these visceral tissues of strain 2 guinea pigs resembled those seen in GPCMV-infected cultured guinea pig cells but differed from those observed in the infected salivary gland duct cells. Strain 2 guinea pigs are a useful animal model for studying disseminated infection in CMV-associated human diseases.     

Bone-marrow interferon

Nucleated bone-marrow cells of mice, rats, guinea pigs, hens, cows, and man are shown to be capable of producing interferon on induction with Newcastle disease virus in vitro. Interferon production by these cells was characterized by high stability. Bone-marrow interferon is not inferior in its activity to interferon obtained by the use of blood leukocytes and spleen cells.     

Porphyrobilinogen biosynthesis from δ-aminolevulinic acid by the viscera of albino rats

Ability to synthesize porphyrobilinogen (PBG) from -aminolevulinic acid (ALA) was determined in homogenates of tissues of the lungs, heart, liver, kidneys, spleen, pancreas, and small intestine of 77 albino rats. All these organs were found to be able to synthesize PBG. Highest ALA dehydratase activity was found in the liver tissue, followed in descending order by the kidneys, lungs, pancreas, small intestine, heart, and spleen. On the addition of a lead solution to the synthesizing system a significant decrease in enzyme activity was observed in the liver tissue, but in kidney tissue its activity was unchanged. On the addition of lead and D-penicillamine simultaneously no changes were found in the toxic effect of lead.M. F. Vladimirskii Moscow Regional Clinical Research Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR S. S. Debov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 12, pp. 687–689, December, 1978.     

Mycoplasma-Associated Induction of Interferon in Ovine Leukocytes

A mycoplasmal species, Acholeplasma laidlawii, isolated as a contaminant from a fetal lamb kidney cell line, was shown to be associated with the induction of interferon in cultures of ovine peripheral blood leukocytes. Broth cultures of the mycoplasma induced between 20 and 230 U of interferon per ml in leukocytes from two adult ewes. The amount of interferon produced correlated with the inoculum size of mycoplasma. Interferon production was associated with replication of the mycoplasma in the leukocyte cultures. Interferon was not induced by sterile mycoplasmal broth, a cell-free filtrate of the mycoplasmal cultures, or heat-inactivated mycoplasmas. The antiviral substance was characterized as interferon by the usual criteria.     

Detection of Als Proteins on the Cell Wall of Candida albicans in Murine Tissues

A murine model of disseminated candidiasis was utilized to determine whether Candida albicans Als proteins are produced in vivo. The kidneys, spleen, heart, liver, and lungs were collected from mice inoculated with one of three C. albicans strains (SC5314, B311, or WO-1). Immunohistochemical analysis of murine tissues by using a rabbit polyclonal anti-Als serum indicated that Als proteins were produced by each C. albicans cell in the tissues examined. Patterns of staining with the anti-Als serum were similar among the C. albicans strains tested. These data indicated that Als protein production was widespread in disseminated candidiasis and that, despite strain differences in ALS gene expression previously noted in vitro, Als protein production in vivo was similar among C. albicans strains. The extensive production of Als proteins in vivo and their presence on the C. albicans cell wall position these proteins well for a role in host-pathogen interaction.     

Distribution of prolactin in selected rat organs and tissues

A single dose of 25 microg prolactin (PRL)/kg of rat body weight was administered to rats subcutaneously. At 1, 2, 3, 4 and 5 h after the injection, selected organs and tissues were taken for analysis. It was found that 1 h after administration, the highest amount of PRL accumulated in the milk (lactiferous) gland, the blood, the ovaries, the pituitary and the liver. Over time, the prolactin content in the selected organs and tissues decreased. PRL is selectively captured by the milk gland, the pituitary, the ovaries, the liver and the heart. Based on the value of the organ or tissue capacity index for PRL, the following order was established for the organs and tissues to which the hormone binds: milk gland > blood > pituitary > ovaries > lungs > liver > cranial bone > spleen > heart > kidneys > muscular tissue > adrenals > adipose tissue > brain.     

Increased expression of matrix metalloproteinases in the murine zymosan-induced multiple organ dysfunction syndrome

Matrix metalloproteinases (MMPs) have been implicated as mediators of tissue damage in several inflammatory diseases. Since the multiple organ dysfunction syndrome (MODS) is thought to result from systemic inflammation, overactivation of MMPs could contribute to the organ damage observed. The expression and activity of several MMPs were studied in a murine model for MODS. Sixty mice were given an aseptic intraperitoneal injection of lipopolysaccharide, followed, after 6 days, by zymosan. At days 2, 5, 8, 12, and 16 after the injection of zymosan, the liver, lungs, spleen, and kidneys were collected from groups of mice for either RNA extraction, gelatinase zymography and collagenase (MMP-1 and -13) assays (six mice per time point), or immunohistochemistry (three mice per time point). A group of nine mice did not receive zymosan and acted as controls. The expression of MMP-2 mRNA in zymosan-treated mice was strongly up-regulated in liver tissue only. For MMP-9, this was the case in all organs examined. Quantitative gelatin zymography demonstrated the near complete absence of any gelatinase activity in tissues from control mice. However, in the liver, lungs, and especially the spleen of zymosan-treated animals, significantly increased activity of proform and active MMP-2 and -9 was observed with time. Overall, MMP-1 and -13 activities were very low in all samples from the liver and lungs. In the spleen, however, high levels of MMP-1 and -13 were observed in zymosan-treated animals. Immunohistochemical staining for MMP-2 was detected in the liver and spleen, but not in lung and kidney tissue of zymosan-treated animals. Staining for MMP-9 could be detected in liver, lung, and spleen tissues of zymosan-treated mice. For both MMPs, staining appeared to be limited to phagocytes. In conclusion, the data suggest a role for MMPs, especially MMP-9, in the pathogenesis of MODS.     

Production of gamma interferon in mice immune to Rickettsia tsutsugamushi.

C3H/He mice immunized by subcutaneous infection with Rickettsia tsutsugamushi Gilliam were examined for the production of immune interferon after intravenous administration of irradiated strain Gilliam antigen, in supernatants of immune lymphocytes stimulated with specific antigen, and after a secondary challenge with viable rickettsiae. Mice administered various doses of irradiated whole-organism antigen 28 days after immunization showed circulating levels of interferon which peaked 4 h after inoculation and were antigen dose dependent. The interferon produced was pH 2 sensitive and stable at 56 degrees C for 1 h and was neutralized by antiserum directed against immune, but not against alpha/beta, interferon. The production of another lymphokine, macrophage migration inhibition factor, paralleled that of interferon. The interferon produced by cultures of spleen cells obtained from immune animals was antigen specific and dose dependent. Peak levels were obtained 48 to 72 h after the addition of antigen. The interferon produced by spleen cell cultures after stimulation with Gilliam antigen was characterized as immune interferon by the same physical and antigenic criteria used for serum interferon. Interferon was produced in vitro by the Thy-1.2+ lymphocyte and required the presence of a spleen-adherent cell population. Immune mice produced high circulating levels of immune interferon after intraperitoneal challenge with viable rickettsiae, which suggested a possible role for interferon in the resistance of immune mice to rechallenge with R. tsutsugamushi.     

Viral replication and interferon production in fetal and adult ovine leukocytes and spleen cells.

Peripheral blood leukocyte and spleen cell cultures derived from adult sheep and from third-trimester (107 to 145 days of gestation) and second-trimester (70 to 98 days of gestation) fetal lambs were examined for their ability to support viral replication and to produce interferon. Bluetongue virus, Herpesvirus hominis type 2, and Chikungunya virus failed to replicate in either leukocyte or spleen cell cultures derived from adult ewes or in cultures from second- or third-trimester fetal lambs. Similarly, peripheral blood leukocytes from adult sheep or third-trimester fetal lambs did not support the replication of Semliki Forest virus, vesicular stomatitis virus, Newcastle disease virus, or vaccinia virus. No major differences were observed in the ability of fetal and adult leukocytes to produce interferon in response to viral infection. In contrast, mean interferon titers induced by bluetongue virus, H. hominis type 2, and Chikungunya virus in spleen cells from second-trimester fetuses were 4- to 10-fold greater than those induced in spleen cells from adult ewes. Variations in interferon levels induced on separate occasions with cells from the same donor age group were observed. The antiviral substance induced in both the fetal and adult cell cultures fulfilled the usual criteria for characterization as interferon.     

The pregnant guinea-pig as a model for studying influenza virus infection in utero: infection of foetal tissues in organ culture and in vivo.

Organ cultures of guinea-pig foetal tissues showed a similar pattern of susceptibility to influenza virus to that already observed for human (Rosztoczy et al., 1975) and ferret (Sweet, Toms and Smith, 1977) foetal tissues. Respiratory, alimentary and urogenital tract tissues were susceptible whereas neural and lymphopoietic tissues were insusceptible. However, of the foetal membranes (amnion, chorion, umbilical cord and placenta) only the chorion was susceptible, in contrast to the corresponding ferret tissues, all of which were susceptible. The insusceptibility of the placenta paralleled that of human placenta which is similarly haemomonochorial in structure. Following intracardial inoculation of high titre virus (ca 10(9-4) EBID50) into pregnant guinea-pigs virus was isolated from all foetal membranes (amnion, chorion, umbilical cord and placenta), but in low titre. Although sporadic isolations were made from foetal tissues (intestine, kidney, heart, liver and spleen) there was no evidence for viral replication in these tissues. These results are discussed in relation to possible infection of the human foetus in utero with influenza virus.