Sequential changes in serum immunoglobulin levels in young RFM mice with host-versus-graft disease.

RFM mice perinatally inoculated with (T6 X RFM)F1 spleen cells develop raidply progressive host-versus-graft (HVG) disease. They are usually dead by 30 days with hyperimmunoglobulinaemia, immune complexes, plasmacytosis and marked T-cell deficiency. In the present studies, sequential quantitative analyses of serum immunoglobulins (Igs) were done, and search was made for high titred antibodies presumed to be major components of the excessive Ig levels. Based on the Ig changes, three stages of disease could be identified. In the first period, which extended from 7 to 10 days, the early appearance of IgA and IgM correlated with previous discoveries of the precocious appearance of germinal centres and enhanced antibody response. The second period, from 10 to 25 days, was characterized by rapid increases in all serum Ig levels in patterns which suggested a variable and selective loss of control of production of the individual Ig classes. IgG1 levels at 25 days averaged twenty-three times the highest adult control value. Failure of IgG2 and IgA levels to surpass adult maxima seemed only to reflect hastened maturation. Lack of success in finding high titred specific antibodies coupled with the previous evidence of poor primary antibody responses suggested the alternative possibility that most of the Igs were non-specific. The third stage, from 25 days to death, saw the apparent decline in IgG1 and IgG2 levels, and the progressive increase in the percentage of HVG mice with IgM levels above the adult maximum. It is proposed that the apparent divergence between Ig and antibody forming capacity is related to the severe disruption of the T-cell system induced by the HVG reaction.     

CD8 serum levels in acute graft-versus-host disease diagnosis

Attempts to identify an early and discriminating marker of acute graft-versus-host disease (aGvHD) have been unsuccessful. The levels of soluble CD4 and soluble CD8 in serum correlate with T cell subset activation and may be important in monitoring and characterizing immunological processes. We determined serum soluble CD4 (sCD4) and sCD8 levels with a two-site sandwich enzyme immunoassay on patients' serum samples collected prior to bone marrow transplantation and weekly after transplantation until day +28. No significant increment of sCD4 was documented in each determination. sCD8 rose significantly before diagnosis or development of maximal clinical symptoms in patients with grade II–III aGvHD than grade 0–I aGvHD [at day +21—median value 447 IU/ml; range 94–713; versus 1136 IU/ml, range 790–1416P=0.002); at day +28—median value 443 IU/ml, range 73–992, versus 1164 IU/ml, range 625–1960P=0.005)]. On the day of marrow infusion the sCD8 levels were significantly higher in patients who subsequently developed grade II–III than in patients with grade 0–I aGvHD (median value 155 IU/ml, range 10–332, versus 350 IU/ml, range 283–830;P=0.003). Careful monitoring of sCD8 is a useful tool for a prompt aGvHD diagnosis and may be used in a clinical bone marrow transplantation setting.     

Enteral human serum immunoglobulin treatment of cryptosporidiosis in mice with severe combined immunodeficiency.

The anti-cryptosporidial immunoglobulin G antibodies in two commercially available human serum immunoglobulin (HSIG) products were quantified and characterized. The mean level of Cryptosporidium parvum-specific immunoglobulin G in HSIG was eightfold higher than the antibody level found in the sera of three immunocompetent individuals convalescing from cryptosporidiosis. However, HSIG products displayed no reactivity to cryptosporidial antigens in immunoblot analyses, while convalescent-phase sera demonstrated characteristic banding patterns. When HSIG was given to newborn severe combined immunodeficiency (scid) mice before and shortly after experimental infection, a decreased intensity of infection was observed in the intestines of the mice compared with that of control mice. However, there was no difference in mortality or histopathologic findings in the intestines of HSIG-treated and control mice when treatment was not started until 22 days of age. These results indicate that HSIG may be beneficial when given prophylactically; however, HSIG cannot eradicate cryptosporidia from mucosal surfaces in an established infection.     

The human X-chromosome and the levels of serum immunoglobulin M

The serum concentrations of immunoglobulins G, A and M were measured in a sample of 93 pairs of monozygotic twins, their spouses, and their offspring. The hypothesis that the human X-chromosome carries genes which control the levels of immunoglobulin M was tested with three different approaches. Our results indicate that environmental factors are primarily responsible for the observed variation in the levels of IgG and IgA. The variance of IgM seems to be mostly the result of X-linked gene effects, with women having higher IgM levels than men.     

Enhanced resistance to Corynebacterium pseudotuberculosis infections associated with reduced serum immunoglobulin levels in levamisole-treated mice.

The effect of levamisole (1-2,3,5,6-tetrahydro-6-imidazo[2,1-b]thiazole monohydrochloride) treatment on the course of a Corynebacterium pseudotuberculosis infection and the ability of either vaccinated or unvaccinated randomly bred mice to resist challenge were investigated. It was found that either a double treatment of levamisole administered with or without a sublethal inoculation of C. pseudotuberculosis or a single treatment of levamisole alone had a significant effect on the ability of mice to resist challenge with virulent C. pseudotuberculosis. This prolonged, enhanced nonspecific and specific resistance was associated with a quantitative reduction in the geometric mean serum immunoglobulin levels, with the immunoglobulin G2 and immunoglobulin A levels being more severely affected. As the enhanced nonspecific resistance to C. pseudotuberculosis was associated with depressed serum immunoglobulin levels in the mice pretreated with levamisole alone, it was suggested that the cell-mediated immunity may play a more important role than the humoral immunity in the resistance to C. pseudotuberculosis infection.     

Age-related serum immunoglobulin E levels in healthy subjects and in patients with allergic disease

Serum IgE levels were measured by a paper-disc radioimmunoassay technique (PRIST) in 425 nonallergic subjects and in 570 patients with asthma, 244 with allergic rhinitis, 48 with asthma and eczema (atopic dermatitis), 49 with eczema but without asthma, and 57 with chronic urticaria. The data are presented for age groups in geometric means and standard deviations and by modal distribution. Normal mean IgE levels for the total sample were 32 IU/ml with highest levels (mean of 51 IU/ml) in school-age children. The highest IgE levels were found in patients with both asthma and eczema (mean of 985 IU/ ml), followed by asthma alone (305 IU/ml), eczema alone (273 IU/ml), and allergic rhinitis (171 IU/ml). The values for our United States population were higher than those reported from Scandinavian countries but lower than those reported from Canada. The geometric mean plus 1 SD (64 IU/ml for infants, 150 IU/ml for schoolchildren, and 100 to 120 IU/ml for all other age groups) appears to be the most useful limit of normalcy. Overlaps with normal values are largest for urticaria, eczema, and allergic rhinitis and least for patients with allergic asthma.     

The relation of serum immunoglobulin A and delayed hypersensitivity skin tests to serum immunoglobulin E levels.

Serum IgA and IgE levels were obtained in 706 patients, 294 of whom were also tested for cutaneous delayed hypersensitivity. The results do not support the concept that a deficiency of serum IgA or of T lymphocyte function underlies the elevated serum IgE levels observed in a majority of allergic patients.     

SENP1对小鼠B细胞数量及血清免疫球蛋白的影响初探

本研究利用Cre/LoxP系统构建B细胞条件性敲除Senp1基因小鼠模型(CKO),探讨了SUMO特异性蛋白酶1(SENP1)对C57BL/6小鼠B细胞数量以及对血清免疫球蛋白产生的作用。利用流式细胞术检测对照组(WT)及实验组(CKO)小鼠骨髓、脾脏及淋巴结B细胞含量及分布情况,结果显示,Senp1敲除后可显著降低脾脏和淋巴结中B细胞的比例,而对骨髓中的B细胞比例没有影响。利用ELISA方法检测血清中免疫球蛋白IgM、IgG、IgG3及IgG1亚型的水平,结果显示CKO小鼠血清IgG1的含量明显低于WT小鼠,而IgM、IgG及IgG3含量没有显著差别。以上结果提示SENP1在小鼠外周淋巴器官B细胞分布及IgG1产生过程中可能起调控作用。     

Changes in immunoglobulin levels in zinc-deficient mice infected with Trypanosoma musculi.

A metabolic imbalance technique was used to study the effects of zinc deficiency on immunoglobulin levels in mice infected with Trypanosoma musculi or immunized with parasite products. Zinc-deficient mice developed higher numbers of parasitemia earlier and exhibited prolonged infection. Irrespective of the diet, higher IgG1, IgG2b, and IgM levels, lower IgG2a and IgA levels, and uniform IgG3 levels were exhibited primarily by mice infected with T musculi and to a lesser extent by mice immunized with parasite products. Zinc-deficient mice showed smaller increases in IgG1 and IgM, but larger gains in IgG2b compared with mice on full-complement and pair-fed diets. However, IgG2a decreased significantly in zinc-deficient mice.     

A case of refractory adult-onset Still's disease with high serum interleukin-18 levels treated with monitoring of serum levels of cyclosporine

We report the case of a 31-year-old woman who developed adult-onset Still's disease (AOSD) with a high level of serum interleukin (IL)-18. Although treated with high dose steroids, she suffered repeated remissions and her condition deteriorated. After we administered oral cyclosporine A (CsA), 200 mg/d, monitoring C2 and trough levels, her symptoms improved significantly. We decreased the dose of methylprednisolone slowly without noting a relapse. The use of CsA accompanied by C2 and trough level monitoring should be considered for refractory AOSD patients with high levels of serum IL-18.     

Bacteriostatic enterochelin-specific immunoglobulin from normal human serum.

Heat-inactivated normal human serum produces iron-reversible bacteriostasis of a number of microorganisms. This inhibitory effect was abolished by adsorption of serum with ultraviolet-killed cells of species that produce the siderophore enterochelin. Bacteriostasis also was alleviated by adsorption of serum with 2,3-dihydroxy-N-benzoyl-L-serine, a degradation product of enterochelin, bound to the insoluble matrix AH-Sepharose 4B. The adsorption process did not add iron or enterochelin to serum, nor did it remove transferrin. The immunoglobulin fraction from normal human serum was isolated; when added to a defined medium (M199) prepared so as to mimic normal human serum, the immunoglobulin rendered the medium inhibitory to an enterochelin-defective strain of Salmonella typhimurium. Adsorption of this medium with AH-Sepharose 4B-2,3-dihydroxy-N-benzoyl-L-serine removed the inhibition. Our results indicate that enterochelin-specific immunoglobulins exist in normal human serum. These immunoglobulins may act synergistically with transferrin to effect bacteriostasis of enterochelin-producing pathogens.     

Rapid subcutaneous immunoglobulin administration every second week results in high and stable serum immunoglobulin G levels in patients with primary antibody deficiencies

Subcutaneous immunoglobulin G (SCIG) infusions as life-long replacement therapy in patients with primary antibody deficiences (PAD) is being applied increasingly. However, only a few published pharmacokinetic studies are available for this route of administration. Therefore, the pharmacokinetics of a 16% immunoglobulin G (IgG) preparation intended for subcutaneous use were investigated in patients with common variable immunodeficiency and X-linked agammaglobulinaemia. SCIG infusions (200 mg/kg body weight) were administered to 12 adult patients every 14 days for 24 weeks (total of 144 infusions). Pharmacokinetic parameters were determined based on serum IgG trough levels and antibody levels against tetanus. The median half-life of the total serum IgG and for the tetanus antibodies was 40.6 and 23.3 days respectively. Median in vivo recovery of serum IgG and tetanus immunoglobulins were 36% and 46% respectively. Median, preinfusion serum IgG trough levels per patient were high without major variations between infusions and ranged from 7.24 to 7.86 g/l. Safety, in terms of adverse events including systemic adverse reactions and local tissue reactions at infusions sites, was monitored throughout the study. Six mild, local tissue reactions were observed during the study in one patient. No systemic adverse reactions related to the study drug were observed and no serious other adverse event occurred during the study. It is concluded that the bi-weekly SCIG therapy was well tolerated in the study and that it results in high and stable serum IgG levels, offering an alternative therapy regimen to patients suffering from PAD.     

An immunohistological study of human hepatic graft-versus-host disease.

Using immunohistological techniques, leucocytes were enumerated in portal tracts and bile duct epithelium in bone marrow transplant recipients with and without evidence of hepatic graft versus host disease (GvHD) and compared with normal subjects. Samples were obtained 8-169 days after transplantation. In marrow recipients without graft versus host disease (GvHD), inducer and suppressor/cytotoxic lymphocytes were reduced in number in the portal tracts compared with normal subjects. In GvHD, suppressor/cytotoxic lymphocytes were increased relative to non-GvHD recipients, but did not exceed normal values, while inducer cell numbers remained low. Natural killer cells (HNK1+) were not found in normal subjects, were present in small numbers in non-GvHD transplant cases and significantly increased in GvHD. The total number of portal tract leucocytes was not elevated in GvHD and the changes in the relative proportions of cells were similar to those that have been observed in the peripheral blood after transplantation. There was no increase in the number of lymphocytes expressing the activation markers Tac, T10 and HLA-DR nor in the number of leucocytes within the bile duct epithelium itself. These findings differ from those we have previously obtained in a similarly treated group of patients with cutaneous GvHD where lymphocytes were increased in the epithelium and stroma and expressed activation markers. Like the epidermis of the skin, however, the bile duct epithelium showed increased staining for HLA-DR antigens in all cases, but focal staining was also present in four of the seven marrow recipients without GvHD. The significance of these findings is discussed.     

Determination of normal human fetal immunoglobulin M levels.

Immunoglobulin M (IgM) levels were measured in 198 cord blood samples from 192 apparently normal pregnancies from 24 weeks of gestation to term. Simple linear regression analysis yielded a standard curve for IgM development during pregnancy showing a 0.5 mg/dl increase in IgM per week of gestation. This curve allows the comparison of fetal IgM levels from pregnancies considered to be at risk for intrauterine infection.     

Determination of immunoglobulin A concentrations in human nasal secretions with a serum immunoglobulin A standard.

By quantitative immunodiffusion tests, nasal secretion immunoglobulin A was underestimated by approximately 5.6-fold when the serum 7S immunoglobulin A standard was employed.     

Reduction of graft-versus-host disease in neonatal F1 hybrid mice.

Pre-immunization of BALB/c (H-2d) mothers with C57BL/10 (H-2b) or CBA/H (H-2k) spleen cells partially protected the F1 hybrid offspring of (BALB/c x C57BL/10) or (BALB/c x CBA/H) matings from graft-versus-host-disease (GVHD) induced by neonatal intraperitoneal inoculation with spleen cells of the paternal strain. The effects achieved were manifest as a reduction in mortality. Experiments to establish whether the phenomenon was antibody mediated were performed by passive pre-immunization of BALB/c mothers with alloantisera obtained from BALB/c previously immunized with C57BL/10 spleen cells. Alloantisera produced an equivalent reduction in GVHD mortality. Some of the F1 mice that survived challenge with paternal strain spleen cells were proven to be haemopoietic chimaeras using immunofluorescence with anti-MHC monoclonal antibodies and polymorphism of the enzyme glucose-phosphate-isomerase present in the strains used. The possible mechanisms of protection from GVHD in our mouse model are discussed.     

The metabolism of different immunoglobulin classes in irradiated mice. V. Contribution of the gut to serum IgA levels in normal and irradiated mice

It was demonstrated in earlier publications that the acute and selective fall in serum IgA levels in irradiated mice was due to a specific failure to renew the plasma pool of this immunoglobulin. In the present work, it is shown that irradiation causes some reduction in the rate of biosynthesis of IgA, but that this phenomenon only partly accounts for the observed disturbance in the metabolism of IgA. This conclusion is based both on direct measurements on the rate of biosynthesis in vitro of IgA by the isolated small bowel wall of irradiated and normal C₃H mice, and on immunohistological studies and plasma cell counts on the gut, spleen and lymph nodes of these animals.On the other hand, it is shown that irradiated mice continue to excrete approximately normal amounts of IgA into their intestinal contents. The conclusion is that irradiation of the gut causes a rechannelling in the output of IgA synthesized by the plasma cells of the lamina propria, the major part being diverted to the intestinal lumen instead of being fed into the circulating pool of serum IgA.