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T-cell activation releases inositol 1,4,5-trisphosphate (IP3), inducing cytoplasmic calcium (Ca2+) influx. In turn, inositol 1,4,5-trisphosphate 3-kinase B (Itpkb) phosphorylates IP3 to negatively regulate and thereby tightly control Ca2+ fluxes that are essential for mature T-cell activation and differentiation and protection from cell death. Itpkb pathway inhibition increases intracellular Ca2+, induces apoptosis of activated T cells, and can control T-cell-mediated autoimmunity. In this study, we employed genetic and pharmacological approaches to inhibit Itpkb signaling as a means of controlling graft-versus-host disease (GVHD). Murine-induced, Itpkb-deleted (Itpkb-/-) T cells attenuated acute GVHD in 2 models without eliminating A20-luciferase B-cell lymphoma graft-versus-leukemia (GVL). A highly potent, selective inhibitor, GNF362, ameliorated acute GVHD without impairing GVL against 2 acute myeloid leukemia lines (MLL-AF9-eGFP and C1498-luciferase). Compared with FK506, GNF362 more selectively deleted donor alloreactive vs nominal antigen-responsive T cells. Consistent with these data and as compared with FK506, GNF362 had favorable acute GVHD and GVL properties against MLL-AF9-eGFP cells. In chronic GVHD preclinical models that have a pathophysiology distinct from acute GVHD, Itpkb-/- donor T cells reduced active chronic GVHD in a multiorgan system model of bronchiolitis obliterans (BO), driven by germinal center reactions and resulting in target organ fibrosis. GNF362 treatment reduced active chronic GVHD in both BO and scleroderma models. Thus, intact Itpkb signaling is essential to drive acute GVHD pathogenesis and sustain active chronic GVHD, pointing toward a novel clinical application to prevent acute or treat chronic GVHD., (© 2020 by The American Society of Hematology.)

Unlabelled: Injection of the LP-BM5 murine leukemia virus into mice causes murine AIDS, a disease characterized by many dysfunctions of immunocompetent cells. To establish whether the disease is characterized by glutathione imbalance, reduced glutathione (GSH) and cysteine were quantified in different organs. A marked redox imbalance, consisting of GSH and/or cysteine depletion, was found in the lymphoid organs, such as the spleen and lymph nodes. Moreover, a significant decrease in cysteine and GSH levels in the pancreas and brain, respectively, was measured at 5 weeks postinfection. The Th2 immune response was predominant at all times investigated, as revealed by the expression of Th1/Th2 cytokines. Furthermore, investigation of the activation status of peritoneal macrophages showed that the expression of genetic markers of alternative activation, namely, Fizz1, Ym1, and Arginase1, was induced. Conversely, expression of inducible nitric oxide synthase, a marker of classical activation of macrophages, was detected only when Th1 cytokines were expressed at high levels. In vitro studies revealed that during the very early phases of infection, GSH depletion and the downregulation of interleukin-12 (IL-12) p40 mRNA were correlated with the dose of LP-BM5 used to infect the macrophages. Treatment of LP-BM5-infected mice with N-(N-acetyl-l-cysteinyl)-S-acetylcysteamine (I-152), an N-acetyl-cysteine supplier, restored GSH/cysteine levels in the organs, reduced the expression of alternatively activated macrophage markers, and increased the level of gamma interferon production, while it decreased the levels of Th2 cytokines, such as IL-4 and IL-5. Our findings thus establish a link between GSH deficiency and Th1/Th2 disequilibrium in LP-BM5 infection and indicate that I-152 can be used to restore the GSH level and a balanced Th1/Th2 response in infected mice., Importance: The first report of an association between Th2 polarization and alteration of the redox state in LP-BM5 infection is presented. Moreover, it provides evidence that LP-BM5 infection causes a decrease in the thiol content of peritoneal macrophages, which can influence IL-12 production. The restoration of GSH levels by GSH-replenishing molecules can represent a new therapeutic avenue to fight this retroviral infection, as it reestablishes the Th1/Th2 balance. Immunotherapy based on the use of pro-GSH molecules would permit LP-BM5 infection and probably all those viral infections characterized by GSH deficiency and a Th1/Th2 imbalance to be more effectively combated., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

It is still unclear whether expanded and activated regulatory T cells (Tregs) in chronic viral infections can influence primary immune responses against superinfections with unrelated viruses. Expanded Tregs found in the spleens of chronically Friend virus (FV)-infected mice decreased murine cytomegalovirus (mCMV)-specific CD8(+) T cell responses during acute mCMV superinfection. This suppression of mCMV-specific T cell immunity was found only in organs with FV-induced Treg expansion. Surprisingly, acute mCMV infection itself did not expand or activate Tregs., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

The gut microbiota has recently been proposed as a novel component in the regulation of host homeostasis and immunity. We have assessed for the first time the role of the gut microbiota in a mouse model of leukemia (transplantation of BaF3 cells containing ectopic expression of Bcr-Abl), characterized at the final stage by a loss of fat mass, muscle atrophy, anorexia and inflammation. The gut microbial 16S rDNA analysis, using PCR-Denaturating Gradient Gel Electrophoresis and quantitative PCR, reveals a dysbiosis and a selective modulation of Lactobacillus spp. (decrease of L. reuteri and L. johnsonii/gasseri in favor of L. murinus/animalis) in the BaF3 mice compared to the controls. The restoration of Lactobacillus species by oral supplementation with L. reuteri 100-23 and L. gasseri 311476 reduced the expression of atrophy markers (Atrogin-1, MuRF1, LC3, Cathepsin L) in the gastrocnemius and in the tibialis, a phenomenon correlated with a decrease of inflammatory cytokines (interleukin-6, monocyte chemoattractant protein-1, interleukin-4, granulocyte colony-stimulating factor, quantified by multiplex immuno-assay). These positive effects are strain- and/or species-specific since L. acidophilus NCFM supplementation does not impact on muscle atrophy markers and systemic inflammation. Altogether, these results suggest that the gut microbiota could constitute a novel therapeutic target in the management of leukemia-associated inflammation and related disorders in the muscle.

The aim of our investigation was to examine the mechanisms of protective features of plaferon LB and fenovit in mice infected with Rausche virus. It was found that in erythroblasts from peripheral blood and spleen, which appeared 12-14 days after infection, the highest percent of determined virus antigens was approximately 47% (range 35,5 to 58,4) which was followed by decrease of the concentration of virus antigens (between 5-12%) in following days and increase of uninfected cells as a result of division of non-contaminated erythroblasts. Mechanism of action of plaferon LB and fenovit was manifested in the reconstitution of mechanisms of apoptosis among erythroblasts, which did not contain synthesized Rausche virus. Impairment of functional activity and antivirus resistance of macrophagal phagocytes are playing an important role not only in the pathogenesis of murine viral leukemia caused by Rausche virus, but also during human leukemia. Correction of functional status of above -- mentioned diseases using plaferon LB and fenovit is presenting as a new and prospective way.

Summary: We previously reported that Moloney murine leukemia virus-ts1-mediated neuronal degeneration in mice is likely a result of both loss of glial support and release of cytokines and neurotoxins from ts1-infected glial cells. Viral infection in some cell types regulates expression of p53 protein, a key regulator of cell proliferation and death. Therefore, we hypothesized that p53 and its dependent genes may be linked with ts1-mediated neuropathology. We examined the presence of p53 and its dependent gene product, a proapoptotic protein bax-alpha, in ts1-induced spongiform encephalomyelopathy. Compared with controls, the lesions of infected animals contained increased levels of p53 and bax-alpha in astrocytes, as shown by strong nuclear p53 and cytoplasmic bax-alpha immunoreactivity in astrocytes. To determine how ts1 affects p53 expression in astrocytes, we then assessed the expression of p53 and its dependent genes, such as bax-alpha and p21, in infected and uninfected immortalized C1 astrocytes and studied possible pathways responsible for p53 accumulation in infected astrocytes. In these studies using mitogen-activated protein kinase inhibitors, infection-induced increases in the p53 level were partially blocked by PD98059, a synthetic inhibitor of MEK1 that is the immediate upstream kinase of extracellular signal-regulated kinases 1/2 (ERK1/2), but not by SB202190, a potent p38 kinase inhibitor. Furthermore, treatment with PD98059 significantly decreased the level of p21 protein, a p53-dependent gene product. These results suggest that ts1 infection may stabilize p53 protein through activation of ERKs in C1 astrocytes, leading to increased expression of the p21 and bax-alpha proteins, both of which induce cell cycle arrest and apoptosis. Our studies suggest that ts1 neuropathology in mice may result from changes in expression and activity of p53, brought about in part by ts1 activation of ERK.

Autoantibodies to alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptors may contribute to chronic hyperexcitability syndromes and neurodegeneration, but their origin is unclear. We examined LP-BM5 murine leukemia virus-infected mice, which manifest excitotoxic brain lesions and hypergammaglobulinemia, for the presence of AMPA-receptor Ab's. Endogenous IgG accumulated upon neurons in the neocortex and caudate/putamen of infected mice and interacted with native and recombinant AMPA-receptor subunits with the following relative abundance: GluR3 > or = GluR1 > GluR2 = GluR4, as determined by immunoprecipitation. In a radioligand assay, IgG preparations from infected mice specifically inhibited [(3)H]AMPA binding to receptors in brain homogenates, an activity that was lost after preadsorbing the IgG preparation to immobilized LP-BM5 virus. These IgGs also evoked currents when applied to hippocampal pyramidal neurons or to damaged cerebellar granule neurons. These currents could be blocked using any of several AMPA receptor antagonists. Thus, anti-AMPA-receptor Ab's can be produced as the result of a virus infection, in part through molecular mimicry. These Ab's may alter neuronal signaling and contribute to the neurodegeneration observed in these mice, actions that may be curtailed by the use of AMPA-receptor antagonists.

Mixed retrovirus infections frequently result in viral pseudotyping in which the genome of a virus is encapsulated within virions possessing a distinct host range from that encoded by the packaged genome. Pseudotyping between different classes of murine retroviruses has been extensively documented in vitro and in vivo and can result in profound changes in viral replication and pathology. A number of non-human retroviruses exhibit infectivity for various heterologous cell lines which often include human cells. Thus there is a possibility of generating heterologous mixed retroviral infections in humans upon the administration of clinical reagents developed in non-human systems. Pseudotyping between retroviruses from heterologous species (e.g., murine and primate retroviruses) has also been documented in mixed in vitro infections, as has pseudotyping between retroviruses and viruses of other families (e.g., rhabdoviruses). The effects of heterologous pseudotyping on viral replication and pathology in vivo have not been extensively studied and represent a potential concern for the development of clinical reagents in heterologous cells.

The bacterial elimination after infection with Listeria monocytogenes was impaired in mice with murine acquired immunodeficiency syndrome (MAIDS) by infection with LP-BM5 murine leukemia virus. Oral administration of hot water extracts of Chlorella vulgaris (CVE) restored the capacity of MAIDS mice to eliminate L. monocytogenes in association with improvement of the deteriorated immune response to L. monocytogenes. DTH response to Listeria in CVE-treated MAIDS mice was significantly higher than that of MAIDS mice after Listeria infection in association with increases in number of CD4+CD8- and CD4-CD8+ alpha beta T-cells in the infected sites. CVE might be effective in the treatment of opportunistic infection in retrovirus-induced immunodeficient patients.

Certain mouse strains, such as AKR and C58, which possess N-tropic, ecotropic murine leukemia virus (MuLV) proviruses and are homozygous at the Fv-1n locus are specifically susceptible to paralytic infection (age-dependent poliomyelitis [ADPM]) by lactate dehydrogenase-elevating virus (LDV). Our results provide an explanation for this genetic linkage and directly prove that ecotropic MuLV infection of spinal cord cells is responsible for rendering anterior horn neurons susceptible to cytocidal LDV infection, which is the cause of the paralytic disease. Northern (RNA) blot hybridization of total tissue RNA and in situ hybridization of tissue sections demonstrated that only mice harboring central nervous system (CNS) cells that expressed ecotropic MuLV were susceptible to ADPM. Our evidence indicates that the ecotropic MuLV RNA is transcribed in CNS cells from ecotropic MuLV proviruses that have been acquired by infection with exogenous ecotropic MuLV, probably during embryogenesis, the time when germ line proviruses in AKR and C58 mice first become activated. In young mice, MuLV RNA-containing cells were found exclusively in white-matter tracts and therefore were glial cells. An increase in the ADPM susceptibility of the mice with advancing age correlated with the presence of an increased number of ecotropic MuLV RNA-containing cells in the spinal cords which, in turn, correlated with an increase in the number of unmethylated proviruses in the DNA extracted from spinal cords. Studies with AKXD recombinant inbred strains showed that possession of a single replication-competent ecotropic MuLV provirus (emv-11) by Fv-1n/n mice was sufficient to result in ecotropic MuLV infection of CNS cells and ADPM susceptibility. In contrast, no ecotropic MuLV RNA-positive cells were present in the CNSs of mice carrying defective ecotropic MuLV proviruses (emv-3 or emv-13) or in which ecotropic MuLV replication was blocked by the Fv-1n/b or Fv-1b/b phenotype. Such mice were resistant to paralytic LDV infection. In utero infection of CE/J mice, which are devoid of any endogenous ecotropic MuLVs, with the infectious clone of emv-11 (AKR-623) resulted in the infection of CNS cells, and the mice became ADPM susceptible, whereas littermates that had not become infected with ecotropic MuLV remained ADPM resistant.

The role of the complement system in the pathogenesis of pulmonary leukostasis in myelocytic leukemia was studied in a rat model. Acute myelocytic leukemia was induced in six Brown-Norway rats, and complement levels were assayed during the course of the disease. Whole complement activity (CH50) and hemolytic activity of C1q, C3, and C4 decreased from day 16 after induction of the leukemia, when the rats developed pulmonary leukostasis. In addition, local complement activation was established in the lung vessels by immunofluorescence microscopy in advanced stages of pulmonary leukostasis. Finally, following systemic activation of the complement system by injection of cobra venom factor (CVF), leukemic rats (n = 6) died of pulmonary leukostasis 4.5 days earlier than did leukemic controls (n = 6). These findings suggest that, in acute myelocytic leukemia in Brown-Norway rats, pulmonary leukostasis is induced by activation of the complement system. This finding could lead to new modes of treatment for a life-threatening complication of leukemia.

Tumor necrosis factor alpha (TNF alpha) levels were determined by enzyme-linked immunosorbent assay (ELISA) and by cell culture bioassay in supernatants of lipopolysaccharide-stimulated feline monocyte cultures and in cat serum samples. There was a good correlation between the results obtained by the two methods. From the fact that TNF alpha was neutralized quantitatively by antibodies to human TNF alpha in feline monocyte supernatants and in feline sera, it was concluded that feline TNF alpha immunologically cross-reacts with human TNF alpha and that the human TNF alpha ELISA can be used to quantitate feline TNF alpha. During the first 6 months after experimental feline immunodeficiency virus (FIV) infection no differences in serum TNF alpha values were observed between infected and non-infected cats. TNF alpha levels increased significantly after primary vaccination with a feline leukemia virus (FeLV) vaccine in FIV infected cats over those in the non-infected controls. During secondary immune response TNF alpha levels rose transiently for a period of a few days in both the FIV positive and the FIV negative cats. After FeLV challenge, TNF alpha levels increased in all animals challenged with virulent FeLV for a period of 3 weeks. This period corresponded to the time necessary to develop persistent FeLV viremia in the control cats. It was concluded from these experiments that in the asymptomatic phase of FIV infection no increased levels of TNF alpha are present, similar to the situation in asymptomatic HIV infected humans. Activation of monocytes/macrophages in FIV infected cats by stimuli such as vaccination or FeLV challenge readily leads to increased levels of TNF alpha.

When neonatal FVB/N mice were inoculated with ts1, a temperature-sensitive mutant of Moloney murine leukemia virus TB, they developed a progressive bilateral hindlimb paralysis and immunodeficiency leading to death 4 to 6 weeks after inoculation. T lymphocytes have been shown to be primarily responsible for this ts1-induced syndrome. Here we compare the role played by each subset of T lymphocytes, i.e., CD4+ and CD8+ T cells, in disease development. Mice were depleted of a specific subset for the first 10 days of their lives by using either anti-CD4 or anti-CD8 monoclonal antibodies in vivo. Disease development in these mice was then monitored. Depletion of CD4+ T cells significantly attenuated the ts1-induced syndrome: virus replication was decreased, disease latency was extended, and death was prevented in 60% of the mice. Similar treatment with anti-CD8 antibody had almost no effect on disease progression. However, when depletion was begun 2 weeks after neonatal ts1 inoculation, CD4+ T cell depletion did not affect disease development. ts1 infected CD4+ and CD8+ T lymphocytes equally well in vivo, as shown by flow cytometric analysis, but virus replication was restricted primarily to the CD4+ subset of T cells, as found by in vitro assay. Hence, CD4+ T lymphocytes play an important role in the development of ts1-induced paralysis and immunodeficiency. The mechanism of this CD4+ T-cell-mediated disease production by ts1 is not clear; however, increased replication of ts1 in the CD4+ T cells, especially in the early stages of the disease, seems to play a crucial role.

The whole skull and spinal column were histologically studied in 55 suckling mice (intact and with transplantable leukemia). It was stated that the lesion of the vertebral canal prevailed due to the extensive growth of leukemic infiltrates in the bodies of the vertebra which had incompletely formed cortical plate of the bone tissue. Such age-dependent unfinished formation of the bone marrow tissue may predispose a rapid development of neuroleukemia.

Most of the B cells from bovine leukemia virus (BLV) infected cows in persistent lymphocytosis (PL) were known to express the CD5 T-cell marker but it was not known whether this peculiar membrane phenotype relates to an activation state. It was demonstrated that these B cells were also flagged by two other membrane markers normally borne by cells belonging to the myeloid lineage (namely CD11b and CD11c). Moreover, cell cycle analysis illustrated that a significant percentage of these B cells (greater than 15%) left their resting (G0/G1) status and progressed through the cell cycle. In addition, T-cell-depleted peripheral blood mononuclear cells from animals in PL were shown to proliferate in response to a IL-2-containing supernatant (MLA 144). These results indicate that the CD5+ B cells from BLV-infected cows in PL are activated cells.

Effects of cocaine administration on lipid peroxidation and liver damage in immunocompromised mice fed different levels of dietary proteins were investigated. Indices of lipid peroxidation and serum aminotransferases as evidence of free radical attack and liver damage were compared in mice fed a low protein (4%) or regular protein diet (20% protein) for 3 weeks and then infected with murine leukemia virus and given daily intraperitoneal injections of increasing progressive doses of 5-45 mg.kg-1.day-1 of cocaine for 11 weeks. Cocaine administration significantly increased hepatic triglycerides, serum aminotransaminases, conjugated dienes, lipid fluorescence, and malondialdehyde levels. These changes were exacerbated by retroviral infection and also by protein undernutrition. Retroviral infection additively increased indices of cocaine-induced lipid peroxidation and hepatic damage. Significant increases in indices of lipid peroxidation and greater liver injury were also detected in similarly treated mice that received the low protein diet compared with well-nourished mice. These results show that immunocompromised mice fed low levels of dietary protein form significantly increased immunogenic lipid peroxidation adducts during cocaine treatment.

The effects of acute and chronic ethanol administration on tumor progression and metastasis were studied in rat models of leukemia and breast cancer, respectively. Acute administration of 1.5-3.5 g of ethanol/kg body weight significantly reduced survival of rats injected with CRNK-16 leukemia cells in a dose-related manner. Acute administration of 2.5-3.5 g of ethanol/kg body weight, one hour before tumor inoculation, or chronic consumption of liquid diet containing ethanol for two weeks before and three weeks after tumor inoculation, significantly increased the number of lung metastases of MADB106 mammary adenocarcinoma. The ethanol-induced increase in the number of metastases was not correlated with plasma levels of corticosterone and was not altered by the opiate antagonist naltrexone. Incubation of spleen cells in vitro in the presence of ethanol, at concentrations comparable to those measured in the blood of ethanol-treated rats, significantly suppressed natural killer (NK) cell activity against MADB106 cells in a standard chromium-release assay and decreased the binding of effector to MADB106 tumor cells. However, neither acute nor chronic ethanol administration in vivo altered splenic NK activity against this tumor in the same in vitro assay, in which the ethanol would have been washed away. These results suggest that, in the presence of ethanol, tumor progression is facilitated. The possibility that this facilitation is related to ethanol-induced impairment of the normal tumoricidal interaction between NK and tumor cells is discussed.

In order to study the role of the spleen in erythropoiesis during AKR/O leukaemogenesis, we have cultured bone marrow and spleen erythroid colony-forming units (CFU-E) and burst-forming units (BFU-E) from AKR/O mice (n = 40) with leukemia of varying severity and type of manifestation. Mice with leukaemia/lymphoma had reduced concentrations of bone marrow CFU-E and BFU-E as compared to healthy, age-matched AKR/O mice. The spleen content of CFU-E was increased, and highest in mice with a spleen size between 500 and 1000 mg. The largest spleens had a somewhat lower CFU-E content. Mice with the highest spleen CFU-E content most often had a normal PCV; however, 4/7 had a normal bone marrow CFU-E concentration. During AKR/O leukaemogenesis with development of spleen enlargement, the spleen may act as an erythropoietic organ, and contribute to maintaining a normal PCV. This may be a temporary ability which is reduced or lost with further progress of the disease.

The pathogenesis of pulmonary leukostasis in leukemia was studied in a rat model by investigating the course of its development. Leukemia was induced by inoculating rats with leukemic cells. The earliest stage of leukostasis was found from day 14 onward, when leukemic cells appeared in the peripheral blood, and was characterized by accumulation of leukemic cells at the capillary level. Simultaneous with the increase of leukemic cell concentrations in the peripheral blood, accumulation in capillaries increased gradually over a period of several days. This was accompanied by increasing severity of tachypnea. Shortly before death, aggregates consisting almost solely of leukemic cells were found in medium-sized blood vessels. This stage was rapidly followed by the end-stage, characterized by complete obstruction of the lung vasculature--including the largest arteries and veins--by leukemic cell aggregates, giving rise to extensive hemorrhages and edema. The end-stage was considered to be the cause of death, which occurred 18-26 days after the inoculation. The histological and ultrastructural findings in this study suggest that besides the size and stiffness of individual leukemic cells, interactions not only between leukemic cells, but also between leukemic cells and the endothelium play a role in the pathogenesis of pulmonary leukostasis.

The ts1 mutant of Moloney murine leukemia virus TB (MoMuLV-TB) causes a degenerative neurologic and immunologic disease in mice characterized by development of spongiform encephalomyelopathy that results in hind-limb paralysis, marked thymic atrophy associated with immunodeficiency, and generalized body wasting. T cells, particularly CD4+ helper T cells, play a key role in the pathogenesis of the disease induced by ts1. Therefore, ts1 is unique among the described murine retroviruses in its ability to afflict both the central nervous system (CNS) and the T-cell compartment of the immune system in the same host. This particular ability to cause degenerative diseases involving both the CNS and immune system is shared by the lentiviruses responsible for development of the acquired immunodeficiency syndromes of humans and macaques. Our goal has been to elucidate the specific cellular and molecular mechanisms that underlie this neuro- and immunopathogenicity of ts1. We have previously reported that the primary neuropathogenic determinant of ts1 maps to a single amino acid substitution, Val-25-Ile, in the precursor envelope protein gPr80env. Further, at the restrictive temperature, the Val-25-Ile substitution did not prevent oligomerization of the gPr80env proteins; however, the structure of the oligomer was incompetent for transport from the ER to the Golgi. These findings suggest that the cytopathic effect of ts1 in neural cells might be due to accumulation of the gPr80env oligomers in the ER. Since glial cells are targets of ts1 infection in vivo, primary astrocytic cultures were established and the cytopathic effect of ts1 and MoMuLV-TB on these cells assessed. Both viruses replicate well in astrocytes and their replication is cytopathic, albeit to different degrees. The ts1 mutant appears to produce greater cell killing than the wild-type virus. Furthermore, it was found that the rate of processing of gPr80env of ts1 in astrocytes is slower than that of MoMuLV-TB. Therefore, the inefficient transport and processing of gPr80env of ts1 appears to correlate with its cytopathic effect in these cells. Electron microscopic studies of the ts1-infected astrocytes revealed large numbers of aberrant particles in the ER. The in vitro cytopathic effect of ts1 on astrocytes may reflect what happens in vivo. An indirect mechanism of neuronal-cell killing by ts1 is proposed.

A total of 147 cats positive for FeLV were retrospectively studied to determine the incidence of ocular disease. Of those cats, 97 had clinical cases of the disease and 50 were artificially infected with the virus. The incidence of ocular disease among FeLV-positive cats with clinical signs of disease was less than 2%, and represented less than 0.1% of the total feline cases for the 5-year period studied. The only ocular findings that could be associated with FeLV were pupillary and motility abnormalities. Retinal hemorrhage and subsequent degeneration found in experimentally infected and naturally infected cats were secondary to profound anemia, which was secondary to FeLV infection. On the basis of the literature and our findings, FeLV is not a major cause of primary or secondary ocular disease in the cat. Anterior uveal disease (iris bombé) was detected in 1 of 147 FeLV-positive cats, and the incidence of secondary infectious disease was zero.

3'-Azido-3'-deoxythymidine (AZT) is used in the management of acquired immunodeficiency syndrome (AIDS) and AIDS-related complex (ARC). The myelotoxic actions of AZT are well known but its effect on platelets is not clear. We studied the effect of AZT at 1 and 2.5 mg/ml in drinking water on platelets in a murine model of AIDS. Three stages of the disease were examined, as determined by the serum IgM levels and other physical features. As early as 15 days after the initiation of drug treatment, AZT was found to significantly increase platelet production. To ascertain that this activity was authentic, a further study was carried out using uninfected mice. Mice were given AZT at both doses for 15 and 30 days. All mice on AZT had significantly increased numbers of platelets. These increases were dose and time dependent. AZT is therefore a potent inducer of thrombocytosis and may be a potential candidate in the treatment of thrombocytopenia in AIDS.

Preexistent feline leukemia virus (FeLV) infection greatly potentiated the severity of the transient primary and chronic secondary stages of feline immunodeficiency virus (FIV) infection. Of 10 FeLV-FIV carrier cats, 5 died of experimentally induced FIV infection, compared with 2 deaths in 10 cats infected only with FeLV and 1 death in 7 cats infected only with FIV. FIV-infected cats with preexistent FeLV infections developed severe depression, anorexia, fever, diarrhea, dehydration, weight loss, and leukopenia 4 to 6 weeks after infection and were moribund within 2 weeks of the onset of signs, whereas cats infected only with FIV developed much milder self-limiting gross and hematologic abnormalities. Pathologic findings in dually infected cats that died were similar to those observed previously in cats dying from uncomplicated primary FIV infection but were much more widespread and severe. Coinfection of asymptomatic FeLV carrier cats with FIV did not increase the levels of FeLV p27 antigen present in their blood over that seen in cats infected with FeLV alone. The amount of proviral FIV DNA was much higher, however, in dually infected cats than in cats infected only with FIV; there was a greater expression of FIV DNA in lymphoid tissues, where the genome was normally detected, and in nonlymphoid tissues, where FIV DNA was not usually found. Dually infedted cats that recovered from the primary stage of FIV infection remained more leukopenic than cats infected with FIV or FeLV alone, and their CD4+/CD8+ T-lymphocyte ratios were inverted. One of these cats developed what was considered to be an opportunistic infection. It was concluded, therefore, that a preexistent FeLV infection in some way enhanced the expression and spread of FIV in the body and increased the severity of both the resulting transient primary and chronic secondary stages of FIV infection. This study also demonstrated the usefulness of the FIV model in studying the role of incidental infectious diseases as cofactors for immunodeficiency-causing lentiviruses.

Signs of disseminated intravascular clotting were observed during the development of BNML myelomonocytic leukaemia in rats, when the peripheral leucocyte count exceeded 20,000/microliters and more than 50% blasts were present in the circulation. BNML cells, harvested from blood and tested in appropriate systems, were found devoid of any procoagulant activity (PCA) even following prolonged in vitro incubation with endotoxin. Thus, it appears that these rat leukaemic cells share the same inability to express PCA which had been previously described in peripheral blood mononuclear cells from normal rats. Conceivably, in this rat model, leucocyte PCA does not represent a major trigger of intravascular coagulation and blood clotting is initiated by other, mainly plasmatic, pathways.

Five sheep were experimentally inoculated with BLV in order to study the humoral immune response in animals infected with bovine leukaemia virus (BLV). During experimental periods of 46 months, 2 sheep died with leukaemia and one sheep showed splenomegaly and proliferation of tumour cells. The other 2 sheep were clinically normal. All of the inoculated sheep developed antiviral antibodies 1 month after inoculation and BLV could be re-isolated in lymphocytes 2 to 3 months after inoculation. Antibody against glycoprotein antigen (gp51) of BLV appeared earlier than the antibody against protein antigen (p24) and antibody titres of the former were higher than those of the latter during the course of the experiment. The complement dependent antibody cytotoxicity test was performed for the detection of antibody against BLV-related cell membrane antigen with 2 different kinds of target cells; FLK cells which are foetal lamb kidney cells chronically infected with BLV and SF-28 cells which are sheep fibroblasts transformed with BLV in vitro. All 5 sheep developed cytotoxic antibodies against both types of cells. In sera from two leukaemic sheep, cytotoxic antibody titres against SF-28 cells gradually decreased 30 months after inoculation and finally became negative one to 3 months before they died of leukaemia. However, these leukaemic sheep persistently produced antibodies against gp51 and p24.

Thrombocytopenia is a consistent feature of murine leukemia L5178Y. To define the mechanism(s) associated with the decrease in platelets, serial thrombokinetic studies were performed on various days after intravenous inoculation of 10(6) L5178Y ascites cells. At the nadir of thrombocytopenia (day 5), the recovery and circulating half-time of transfused normal 51Cr-platelets was only one half of control values. The loss of circulating radioactivity (70%) at 1 hr was accounted for by splenic (40%) and hepatic (30%) accumulation of labeled platelets. However, the liver contained three times more radioactivity per milligram of tissue than the spleen. There was no increase in hepatic accumulation of 59Fe-labeled red cells under the same conditions. When 51Cr-labeled leukemic platelets (day 3) were infused into normal animals, the circulating half-time was 50% of control. Despite spleen and liver enlargement, the blood volume of the leukemic mice did not increase. The megakaryocyte concentration remained unchanged after inoculation of leukemic cells, but the diameter of megakaryocytes increased significantly from days 5 to 10. These studies show that thrombocytopenia in mice transplanted with L5178Y leukemia occurs as a result of shortened platelet survival and increased organ sequestration. The increase in hepatic platelet accumulation suggests that the mechanism of thrombocytopenia is platelet specific and is not due to passive organ pooling.

Mice infected with 2 x 10(3) plaque-forming units of mouse cytomegalovirus (MCMV) 3 days after receiving 300 to 400 spleen focus-forming units of Friend leukemia virus developed a more severe MCMV infection than did normal animals. Increased severity was demonstrated by the increased amounts of MCMV recoverable from the salivary glands of leukemic mice 1 to 5 weeks postinfection. In addition, the difference in the number of virus isolations from the kidneys, spleens, livers, and lungs of animals (74 to 120) coinfected with MCMV and Friend leukemia virus compared with animals (49 of 120) infected with MCMV alone was significant (P less than 0.01). Both the 50% lethal dose and 50% infectious dose of MCMV in leukemic mice were lower than in normal animals. MCMV and Friend leukemia virus appear to interact by suppressing the ability of infected spleen cells to respond to mitogen-induced stimulation. The observations of increased severity of MCMV infections in leukemic mice closely parallel the situation observed in human leukemia patients who are at an increased risk of disease due to human cytomegalovirus infections. This mouse model may be useful in assessing the effect of antiviral (cytomegalovirus) therapy.

Six cattle persistently infected with bovine virus diarrhoea virus (BVDV) and seronegative, and two control, virus negative seropositive cattle were inoculated with lymphocytes infected with bovine leukosis virus (BLV). The two controls produced a normal immune response to BLV, developing antibodies at four and five weeks after inoculation. Two of the six cattle persistently infected with BVDV developed a strong antibody response by six weeks after inoculation with BLV. Four developed a depressed response to BLV, characterised in three by a 'hooking' reaction in the immunodiffusion test which persisted in successive bleedings but was interspersed occasionally by a weak positive reaction. In one of these animals, a series of 'hooking' reactions was followed by a number of negative results. The fourth animal remained serologically negative until 16 weeks after inoculation when a 'hooking' reaction was observed followed by a series of negative results. BLV was isolated from all the cattle persistently infected with BVDV at 42 or 58 weeks after inoculation regardless of whether the serum samples gave negative, 'hooking', weak positive or positive reactions in the immunodiffusion test. BLV was consistently isolated from the nasal secretions of a steer which was BVDV negative but seropositive. The possibility of decreased immune responsiveness to BLV in animals persistently infected with BVDV should be considered when formulating regulations governing the testing of animals for freedom from BLV.

An investigation was carried out on 59 mice of high leukemic line C-58. It is shown that aseptic inflammation inhibited the leukemic process in mice with transplantable leukemia, stimulated the myeloid sprout of hemopoiesis and suppressed the lymphoid one, and also decreased the spleen and thymus mass.

M. arthritidis is known to enhance the leukemogenic effect of Rauscher leukemia virus in mice of different strains. As we have found earlier, M. arthritidis enhanced endogenous colony formation in irradiated mice. In this paper we made an attempt to establish a connection between these two phenomena. The mixed inoculation of BALB/c mice with M. arthritidis and Rauscher leukemia virus produced a great increase in the number of endogenous colonies as compared with the effect of mycoplasmas alone, while Rauscher leukemia virus did not enhance the number of colonies. When injected into mice a day before their lethal irradiation, M. arthritidis was shown to increase the survival of mice within 30 days following irradiation. We tried to determine the possible mechanism of the effect of M. arthritidis on hemopoiesis. Our findings led us to the conclusion that the mycoplasmas had no influence on the splenic microenvironment and did not increase the migration of the surviving stem cells to the spleen. It is conceivable that M. arthritidis acts not on the pluripotent stem cells, but rather on the committed precursor cells. If so, the mechanism responsible for the enhancing effect of mycoplasmas on the development of Rauscher leukemia and endogenous colony formation resides possibly in the capacity of M. arthritidis to increase the number and/or sensitivity of target cells to the virus.

Sixteen feline leukemia virus (FeLV)-infected cats with lymphosarcoma (LSA) were treated by extracorporeal immunoadsorption using staphylococcal protein A columns in order to remove immunoglobulin G (IgG) and circulating immune complexes (CIC) from plasma. Complete viral clearance and long-lasting tumor regression were achieved in nine of the cats and tumor regression without virus clearance was observed in two other cats. Since LSA cats rarely go into spontaneous remission, and since other forms of therapy are ineffective, these cats offered a unique system for analyzing details of the immune response to LSA and FeLV as they are cleared. Immunological parameters associated with the FeLV and LSA responses were assessed in detail in three responder cats and three nonresponders during the treatment and follow-up periods. Two serological parameters that always correlated with complete clearance of LSA were development of precipitating antibodies against FeLV-C gp70 and development of cytotoxic antibodies that kill cultured FL74 LSA cells in the presence of complement. The precipitating antibodies were detected prior to the clearance of LSA and prior to the detection of free cytotoxic antibodies. One serological parameter that always correlated with complete clearance of. FeLV was development of free antibodies to FeLV-AB gp70. Quantitative levels of FeLV-specific CIC and feline oncornavirus-associated cell membrane antigen (FOCMA)-specific CIC correlated well with fluctuating levels of the corresponding antigens and antibodies. These results suggest that the staphylococcal protein A treatment columns remove CIC "blocking factors" directly or indirectly and thereby stimulate existing antibody responses. These antibodies mediate clearance of FeLV and LSA.

A study was made of the content of ubiquinone, vitamins A, E, ascorbic, dehydroascorbic and diketogulonic acids (DKGA), and malonic dialdehyde (MDA) in the liver, of the content of glutathione, the activity of superoxide dismutase (SOD) and glutathione reductase in red blood cells, of the content of vitamins A, E and ubiquinone in the spleen of C57Bl/6jG mice with inoculated leukemia La. It was found that in red blood cells of the animals with leukemia, the content of vitamin E and DKGA reduced, the MDA level increased, and the content of glutathione dropped whereas SOD activity rose. Application of the antioxidant complex of vitamins A, E, C appreciably improved the characteristics of enzymatic and non-enzymatic antioxidant protection of the liver and red blood cells of the leukemic animals without exerting any noticeable effect on the content of vitamin E and ubiquinone in the leukemic spleen tissue.

A group of temperature sensitive mutants of Moloney murine leukemia virus (MoMuLV) designated as ts1, ts7, and ts11 rapidly and invariably induce hind-limb paralysis ranging from 28 to 52 days postinjection of neonatal CFW/D mice. These temperature-sensitive mutants are defective in the processing of the precursor of the env protein, gPr80env in infected cells, resulting in the accumulation of gPr80env in the infected cell and production of virions with reduced amounts of gp70, p15E, and p12E when compared to that of the wild-type virion. In contrast, two nonparalytogenic ts mutants, ts3 and ts10, like the wild-type virus, show normal processing of gPr80env in infected cells and production of virions with a similar amount of env proteins to that of the wild-type virion.

Extensive animal studies and clinical observations support an immunosuppressive role for certain antibodies and circulating immune complexes (CIC) in malignant and autoimmune diseases. Investigators have attempted to correct or modulate dysfunction by removal of antibodies or CIC from plasma. Extra-corporeal immunoadsorption of plasma over columns containing a silica matrix and covalently attached highly purified staphylococcal protein A (PROSORBA column) is a procedure that specifically removes those plasma components by the interaction of protein A with the Fc region of IgG. The interaction of CIC with the Fc receptor on protein A has three specific results. First, there is direct removal of immunosuppressive CIC from the circulation. Studies of CIC-mediated immunosuppression in experimental systems have shown dose-response relationships over wide ranges of CIC concentrations. Thus, removal of CIC relative to the IgG antibody may be expected to exert some stimulation of the immune system. Second, the complement system is activated. Elevated levels of C3a, C4a, and C5a are observed in patients' circulating plasma after PROSORBA treatment. These levels peak one to three hours post-perfusion and are near normal levels by six hours post-perfusion. These complement components are stimulators of growth and activity of immune cells. In addition, by binding to CIC they stimulate clearance of CIC by the reticuloendothelial system. Thus, treatments may induce removal of more CIC than could be anticipated by the binding capacity of treatment columns. Third, antibody is released from CIC. Interaction of CIC with bound protein A with or without the aid of activated complement components leads to liberation of free antibody. Depending upon other factors, eg, amount of circulating antigen and/or unbound IgG, either free antibody or CIC containing more antibody relative to antigen (or both) may be infused into patients with the posttreatment plasma. Such CIC function as immune stimulators rather than suppressors of immune cell activity. The consequences of the treatments are summarized as follows. Stimulation of immune cellular activity is seen one to three hours posttreatment. During the first one to three treatments, cells of the granulocyte/macrophage series show the greatest increase. During and after treatments 2 to 4, lymphocytes show the greatest increase. At this point, increased blastogenic response to mitogens is observed along with an increase in the T helper/suppressor cell ratio.(ABSTRACT TRUNCATED AT 400 WORDS)

We compared the incidence of lymphomas induced by Gross leukemia virus (GLV) between spontaneously hypertensive rats (SHR) with a congenital T-cell depression related to thymic dysfunction and normal Wistar rats, the original strain of SHR. Of 20 SHR given neonatal injections of GLV, only 3 (15%) died with thymic lymphomas about 100 days after the virus infection. In contrast, 27 of 28 Wistar rats (96%) developed lymphomas of mostly thymic origin. The 3 lymphomas derived from the SHR bore only a Thy 1.1 antigen, whereas most of the lymphomas derived from Wistar rats carried not only a Thy 1.1 antigen but also a guinea pig red blood cell rosette receptor and a T (W3/13) antigen. Grafts of 1-week-old male Wistar thymus into the neonatal female SHR promoted a differentiation of thymocytes and markedly increased the incidence of the lymphomas which were positive for a guinea pig red blood cell rosette receptor and a T-antigen; grafts of 1-week-old SHR thymus, however, failed to do this. These results suggest that the low incidence of GLV-induced lymphomas in SHR may correlate closely with the absence or decreased numbers of the rosette-forming thymocytes which are presumably the target cells for GLV.

Since mice can synthesize ascorbic acid but man cannot, the ascorbate status in murine and human leukaemia was compared. The decline in plasma ascorbate concentration in both cases indicates that vitamin C deficiency occurs in malignancy. Analysis of tissue ascorbate values in mice also indicated that an enhanced rate of utilization of this vitamin occurs during malignancy, as does an increased rate of excretion, and both events may be responsible for vitamin C deficiency. The hepatic ascorbate values suggest an endeavour by the animals to compensate for the loss through increased synthesis and storage of the vitamin, at least in the early stages of the disease.

Friend murine leukemia virus complex (FV)-induced immunosuppression was studied by assaying splenic anti-SRBC PFC responses and plasma antibody titers in mice at various times after FV inoculation. Genes located within the H-2 complex were found to influence resistance to FV-induced immunosuppression. Near normal responses were observed in mice having the H-2a/b or H-2b/b genotype, whereas mice having the H-2a/a genotype were suppressed. This H-2 effect was observed not only in mice having heterozygous C57BL/10 X A background genes, including Rfv-3r/s, but also was apparent in mice having homozygous A-strain background genes, including Rfv-3s/s. Therefore, the Rfv-3 gene did not appear to convey resistance to FV-induced immunosuppression. The suppression in susceptible H-2a/a mice was characterized by a partial suppression of the IgM response and a profound suppression of both the primary and secondary IgG responses. Neither splenomegaly nor viremia alone appeared to be sufficient for the induction or maintenance of the immunosuppression. The mechanism of suppression was unclear, but both B lymphocyte and T lymphocyte functions appeared to be altered.