Loss of B Cells in Patients with Heterozygous Mutations in IKAROS.

BACKGROUND: Common variable immunodeficiency (CVID) is characterized by late-onset hypogammaglobulinemia in the absence of predisposing factors. The genetic cause is unknown in the majority of cases, and less than 10% of patients have a family history of the disease. Most patients have normal numbers of B cells but lack plasma cells. METHODS: We used whole-exome sequencing and array-based comparative genomic hybridization to evaluate a subset of patients with CVID and low B-cell numbers. Mutant proteins were analyzed for DNA binding with the use of an electrophoretic mobility-shift assay (EMSA) and confocal microscopy. Flow cytometry was used to analyze peripheral-blood lymphocytes and bone marrow aspirates. RESULTS: Six different heterozygous mutations in IKZF1, the gene encoding the transcription factor IKAROS, were identified in 29 persons from six families. In two families, the mutation was a de novo event in the proband. All the mutations, four amino acid substitutions, an intragenic deletion, and a 4.7-Mb multigene deletion involved the DNA-binding domain of IKAROS. The proteins bearing missense mutations failed to bind target DNA sequences on EMSA and confocal microscopy; however, they did not inhibit the binding of wild-type IKAROS. Studies in family members showed progressive loss of B cells and serum immunoglobulins. Bone marrow aspirates in two patients had markedly decreased early B-cell precursors, but plasma cells were present. Acute lymphoblastic leukemia developed in 2 of the 29 patients. CONCLUSIONS: Heterozygous mutations in the transcription factor IKAROS caused an autosomal dominant form of CVID that is associated with a striking decrease in B-cell numbers. (Funded by the National Institutes of Health and others.).

Effect of neonatal thymectomy and antilymphocyte globulin on non-specific mitogen stimulation in Holstein-Friesian calves.

Nine of seventeen neonatal Holstein-Friesian calves were thymectomized, treated with antilymphocyte globulin, and monitored for immunologic functional ability for 4 to 6 months. The thymus weights for 4 to 10-day-old calves and 4 to 6-month-old calves indicated a continued increase in total weight. This indicated significant thymic involution had not occurred at 4 to 6 months. Following thymectomy a wasting syndrome was not observed although an increased incidence of a lowly virulent virus infection did occur. A significant decrease in circulating lymphocytes was observed. Peripheral blood lymphocytes were stimulated in vitro by non-specific mitogens, phytohemagglutinin, bacterial lipopolysaccharide and pokeweed mitogen using the whole blood culture method. Observations included a greater response to phytohemagglutinin and pokeweed mitogen in summer months and variable age related response to all mitogens. Lipopolysaccharide stimulation results were inconclusive. It was concluded that neonatal thymectomy was not a satisfactory experimental procedure for the production of selective immunosuppression in the bovine species.

The role of lysosomes in the pathogenesis of copper-induced hepatotoxicity: morphological studies.

Copper-induced hepatotoxicity was evaluated in beige and conventional mice which were injected intraperitoneally with aqueous copper chloride (8 mg per kg body weight per day). Hepatic lesions developed more quickly and were more severe in beige mice and consisted of karyomegaly, progressive accumulation of lipofuscin within hepatocytes and Kupffer cells, acute necrosis of random hepatocytes, and aggregation of leukocytes within the sinusoids. In both groups of mice the hepatic lesion had a centrilobular distribution and closely paralleled the appearance and distribution of copper within hepatocytes. Overall, beige mice had more severe copper-mediated hepatotoxicity, suggesting that normal lysosomes have protective effects for hepatocytes by sequestering copper from other cytoplasmic constituents.

Effects of estradiol on hematopoietic and marrow adherent cells of dogs.

Hematopoietic and marrow adherent cells were examined in 5 ovariohysterectomized dogs treated once with 1.5 mg of 17 beta-estradiol cyclopentylpropionate/kg of body weight. All dogs initially developed thrombocytopenia and neutrophilic leukocytosis, with megakaryocytopenia and an increased myeloid/erythroid ratio of the marrow. Later, 2 dogs became thrombocytopenic and neutropenic, with megakaryocytopenia, decreased myeloid/erythroid ratio, and increased concentration of plasma cells and mast cells in the marrow. The concentration of marrow granulocyte-macrophage colony-forming units (CFU-GM) was decreased at 2 and 3 weeks in all dogs after estradiol treatment. The concentration of marrow fibroblast CFU-F was decreased in all estradiol-treated dogs at 1 and 2 weeks. The ability of marrow adherent cells to support CFU-GM colony formation was unchanged in estradiol-treated dogs. Estradiol sulfate at final concentrations of 10(-8) to 10(-4)M did not affect the in vitro growth of CFU-GM and CFU-F, or the hematopoietic ability of marrow adherent cells from nontreated dogs.

Acute canine ehrlichiosis: platelet survival and factor 3 assay.

Ten dogs experimentally inoculated with Ehrlichia canis were thrombocytopenic 14 days after inoculation. Circulating platelet concentrations had declined rapidly by postinoculation day 10, but remained at or slightly below the 14-day concentration during the remainder of the 42-day experimental period. The percentage of circulating megathrombocytes also increased to 8.7 times preinoculation concentrations indicating the existence of accelerated thrombocytopoiesis. The release of platelet factor 3 (PF-3) from normal canine platelets was delayed after incubation of the platelets in globulin fractions from infected dogs. Inhibition of PF-3 release was attributed to the effect of an unidentified substance present only in the dialyzed globulin fraction of infected canine serums. The presence of antiplatelet antibody in these dogs was not evident. Survival of 51Cr-labeled platelets was decreased in infected dogs, but was most abbreviated in infected dogs that were thrombocytopenic when injected with labeled platelets. Thus, accelerated nonantibody-mediated destruction of platelets contributes markedly to the development of thrombocytopenia in dogs with acute ehrlichiosis.

Effects of cyclophosphamide on the lymphoid tissues and humoral and cellular immune responsiveness of young calves.

Cyclophosphamide (CY) was given IV to 5-month-old calves (ten doses; each dose of 5.0 mg/kg, 2-day intervals between doses). The effects of CY on circulating leukocytes, lymphoid tissues, and the humoral and cellular immune responses were assessed. The numbers of total leukocytes, lymphocytes, and neutrophils and platelets decreased significantly. The lymphocyte population was depleted in the cortex of the thymus and B-dependent areas of the spleen and lymph nodes. Significant decreases occurred in the frequency of the peripheral blood lymphocytes-bearing surface immunoglobulin (Ig) and in serum IgM and IgG concentrations. Primary serum antibody responses to avian erythrocytes and Brucella abortus strain 19 antigens were diminished or delayed. The blastogenic responses of peripheral blood lymphocytes to phytohemagglutinin P, concanavalin A, pokeweed mitogen, and to purified protein derivative and B abortus antigens were enhanced as was the delayed hypersensitivity reaction to the tuberculin skin test. While a diminished humoral immune response was associated with CY treatment, the cell-mediated response was potentiated. The effect of CY was transitory with most variables returning to near base line within 24 days after CY was ceased.

Response to neonatally thymectomized Holstein calves to bacille Calmette Guérin.

Neonatally thymectomized and intact Holstein calves were inoculated intradermally with bacille Calmette Guérin mycobacteria at 4 to 6 months of age. The immune response of the calves was evaluated by in vitro lymphocyte stimulation, with a purified protein derivative of Mycobacterium bovis, and by a cervical skin test. Eight of te calves had been previously infected at 7 to 15 weeks of age with bovine viral diarrhea virus. The cutaneous and in vitro immune responses were not different in neonatally thymectomized calves, whether or not they had been previously infected with bovine viral diarrhea virus. However, a prominent histologic difference in the nature of the primary granulomatous lesions was observed in the thymectomized calves. These injection sites were characterized by few lymphocytes and an increased number of multinucleated giant cells. These observations were interpreted as manifestations of a deficient thymus-dependent effector response in the thymectomized calves.

Serum complement and blood platelet adhesiveness in acute canine ehrlichiosis.

Three dogs inoculated with Ehrlichia canis served as infected controls (group I), 3 dogs treated with cobra venom factor (CVF) after inoculation with E canis were the principals (group II), 3 noninfected dogs were the noninfected controls (group III), and 1 noninfected dog given CVF was the noninfected, decomplemented control (group IV). Thrombocytopenia occurred in all E. canis-inoculated dogs (groups I and II) within 3 weeks after they were inoculated. However, the severity of the thrombocytopenia was moderated in the group II dogs (decomplemented E canis-inoculated). Hypocomplementemia occurred in the CVF-treated dogs (groups II and IV) immediately and persisted for up to 5 days after administration of CVF. Hypocomplementamia also occurred in all E canis-infected dogs (groups I and II) during the period of maximal thrombocytopenia. The mean platelet retention was significantly decreased in all E canis-inoculated dogs and in the CVF-control dog (groups I, II, and IV). There was no correlation between platelet retention and the number of platelets in group III (nontreated, noninoculated) controls. The correlation between the decrease in platelet retention and the number of platelets was significant in the complement-intact infected dogs, yet was not significant in the decomplemented infected dogs. Data indicated that complement consumption occurred in acute canine ehrlichiosis and coincided, to a degree, with the development of thrombocytopenia. Decomplementation of inoculated dogs moderated the severity of the thrombocytopenia. Platelet function is also altered in acute stages of the disease.

Effect of incorporation of serum from dogs with renal impairment on canine erythroid bone marrow cultures.

Serum from dogs with surgically induced renal impairment was incorporated into the medium for erythroid bone marrow cultures. A significant correlation was found between serum activities of erythropoietin and numbers of erythroid colony-forming units grown in culture. Serum creatinine concentrations had no correlation, and serum parathyroid hormone activities had a negative correlation with numbers of erythroid colony-forming units that was below the level of significance. Purified 1-84 parathyroid hormone added to bone marrow cultures was found to be stimulatory to erythroid colony-forming unit growth in higher concentrations, but decreased the number of burst-forming units. Unmeasured substances in the canine serum appeared to have a greater effect on the canine erythroid bone marrow cultures than did creatinine or parathyroid hormone values.

Serum immunoglobulin concentrations of cattle in a herd with bovine leukosis.

Consistent relationship was not found between the serum concentrations of immunoglobulin IgM and IgG and the concentration of blood lymphocytes (Bendixen index) in 289 Holstein-Friesian heifers and cows from a dairy herd in which bovine lymphoma had been identified repeatedly. Serum IgM was present in all cattle, and those cows with highest concentrations of blood lymphocytes generally had higher levels of IgM.

Role of parathyroid hormone in the anemia of chronic terminal renal dysfunction in dogs.

Terminal renal dysfunction (TRD) was induced in 2 groups of dogs by partial surgical ablation of the kidney. Dogs of a control group and of 1 of the TRD groups were maintained on a diet containing normal phosphorus concentration, whereas dogs of the other TRD group were maintained on a low-phosphorus diet. Mild anemia developed in dogs of both TRD groups and could not be attributed to iron deficiency, increased erythrocyte concentration of 2,3-diphosphoglycerate, or absolute deficiency of erythropoietin (EP). Subsequently, all dogs were acutely depleted of approximately 25% of their blood volume. Erythropoietin concentration in dogs of the TRD groups was lower than that of controls, however, erythroid regenerative capacity was comparable with that of control dogs when plasma parathyroid hormone (PTH) concentration was lowered by reduced dietary intake of phosphorus. The PCV in dogs of the chronic TRD groups had a slight positive correlation with serum EP concentration, and a significant (P less than 0.05) negative correlation with plasma PTH and serum phosphorus and creatinine concentrations, using a correlation matrix. There was no longer a significant correlation between plasma PTH concentration and PCV after controlling for serum creatinine concentration by use of a multiple linear regression analysis. A significant (P less than 0.05) negative correlation also was observed between plasma PTH and serum EP concentrations, but not between serum EP and phosphorus or creatinine concentrations. Significance of the EP and PTH association was reduced when analyzed, using a multiple linear regression analysis that included serum creatinine values.(ABSTRACT TRUNCATED AT 250 WORDS)