Multiple myeloma presenting with widespread osteosclerotic lesions.

Sclerotic lesions are rare in malignant monoclonal gammopathies, although they are occasionally associated with POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy and skin changes). In most cases, osseous lesions in POEMS syndrome present as an isolated sclerotic deposit or a combination of both lytic and sclerotic lesions. Diffuse osteosclerosis is extremely rare and may lead to the diagnosis of multiple myeloma, classically known to present as lytic lesions in the skeleton, with or without diffuse osteoporosis. We report a 74-year-old woman with widespread and substantial osteosclerotic lesions, associated with IgA-lambda myeloma, and with no other criteria for POEMS syndrome, and who was rapidly diagnosed with compression of the spinal cord. Detailed knowledge of imaging features in myeloma emphasises the need to consider plasma cell neoplasm in the differential diagnosis of any pattern of bone sclerosis. Although exceptional, multiple myeloma must be borne in mind in the presence of diffuse bone sclerosis.

[The identification of antigens of systems AB0, GM, and haptoglobin (HP) phenotypes in mixed bloodstains of human and domestic goose]. / Identifikatsiia antigenov sistem AB0, GM and fenotipov gaptoglobina (HP) v smeshannykh piatnakh krovi cheloveka i domashnikh gusei

The traditional methods of investigations according to systems AB0, Gm and Hp were used to define the serological specificity of home gooses' blood. The experimental examinations' results related with mixed bloodstains of man and home gooses are described. A possibility is demonstrated to identify the blood group factors of man in bloodstains with admixture of home-goose blood.

Anti-HPA-5b-induced neonatal alloimmune thrombocytopenia: antibody titre as a predictor. Collaborative Study Group.

Anti-HPA-5b is the most commonly found platelet-specific antibody among pregnant women, but it does not cause severe fetal-neonatal alloimmune thrombocytopenia in the majority of affected infants. However, as the sequelae of the affected children may become severe, it is necessary to identify the risk factors for neonatal alloimmune thrombocytopenia. Of 21 354 consecutive pregnant women, 138 [0.65%; 95% confidence interval (CI) 0.54-0.75%], corresponding to 13.2% of the 1049 HPA-5b- women calculated by the gene frequency, were positive for anti-HPA-5b at the first trimester. Anti-HPA-5b was titrated in specimens obtained at the third trimester and antibody-positive women and their neonates were HPA-5 genotyped. Platelet counts in cord blood and 3 d after birth were assessed in the infants born to these mothers. Chi-square analysis showed no significant relationship between the titres of maternal antibody to HPA-5b and the number of pregnancies. There was a significant difference in platelet counts at d 3 between neonates who were compatible (267 x 109/l) or incompatible (220 x 109/l, P < 0.05) with maternal anti-HPA-5b. HPA-5b antibody titres >/= 64 were related to the development of thrombocytopenia (< 150 x 109/l) in neonates 1 d and 3 d after birth. A high titre (>/= 64) had a positive predictive value of 50% for thrombocytopenia 3 d after birth when the infant was HPA-5b+ and a negative predictive value of 100%. These results indicate that a high titre (>/= 64) of anti-HPA-5b is associated with a higher risk of neonatal thrombocytopenia, even if anti-HPA-5b-induced severe thrombocytopenia rarely develops.

Bovine IgG2a antibodies to Haemophilus somnus and allotype expression.

Bovine IgG2a has been implicated in protection against pyogenic infections, including those caused by Haemophilus somnus. To further investigate the role of IgG2a in defense against H. somnus, IgG1 and IgG2a antibodies were purified from antiserum against an immunodominant 40 kDa outer membrane protein (p40) of H. somnus, which was previously shown to passively protect calves against H. somnus pneumonia. The passive protective capacity of anti-p40 IgG1 or IgG2a was evaluated in vivo in calves. Purified anti-p40 IgG1 or IgG2a was incubated with H. somnus for 15 min before intrabronchial inoculation of calves. Bacteria incubated with anti-p40 IgG1 or IgG2a were inoculated into one caudal lung lobe and bacteria incubated with IgG1 or IgG2a from the respective preimmunization serum were inoculated into the contralateral lobe. The volumes of pneumonia in the right and left lungs were determined 24 h later. The difference in volume of pneumonia with H. somnus preincubated in IgG1 pre- and postimmunization anti p40 was less (16 cm3, P = 0.298) than the difference in volume of pneumonia with H. somnus preincubated in IgG2a pre- and postimmunization anti p40 (30 cm3, P = 0.146). Although the differences in lesion size between pre- and postimmunization serum were not statistically significant, the trend suggests IgG2a may be more protective than IgG1. To examine this further, the peptide specificity of these IgG1 and IgG2a antibodies to p40 was examined. After limited proteolysis of p40, IgG2a antibodies reacted with 2 peptides not recognized by IgG1 antibodies. Other peptides were recognized by both isotypes. Since these studies suggested that IgG2a may be important in protection against infection, we then investigated some aspects of the role of the 2 IgG2a allotypes, A1 and A2. In retrospective studies of age differences in expression of IgG2a allotypes, no heterozygotes were detected in calves of 60 d old or less, and fewer heterozygotes were detected in calves 61-120 d old than in cattle older than 270 d (P < 0.01). In a subsequent prospective study of the time course of allotype expression, Holstein calves shown to be heterozygotes expressed the IgG2aA1 allotype early but the IgG2aA2 allotype was not usually detected until 3 to 4 mo of age. Thus, both the retrospective and the prospective studies showed age related differences in expression of the IgG2aA1 and A2 allotypes. This could have implication in protection.

Sequence changes at the V-D junction of the VH1 heavy chain of anti-phosphocholine antibodies alter binding to and protection against Streptococcus pneumoniae.

X-linked immune deficient (Xid) mice fail to produce anti-phosphocholine (PC) antibodies even after immunization with Streptococcus pneumoniae. Consequently, Xid mice are extremely susceptible to infection with S. pneumoniae, PC-specific B cells appear to undergo clonal deletion in Xid mice; however, a new thymus-dependent form of PC, 6-(O-phosphocholine)hydroxyhexanoate (EPC), can rescue PC-specific B cells from the bone marrow presumably by providing T cell help before clonal deletion. Analysis of PC-specific IgG hybridomas from Xid mice revealed utilization of several V-D junctional variants of the VH1 gene segment rearranged to different D and JH gene segments. The majority of Xid anti-PC antibodies exhibit an Asp-->Gly95H replacement at the V-D junction. These Gly95H VH1 variants associate with kappa 1C L chains to produce anti-PC antibodies that: (1) have low relative affinity for PC, (ii) are heteroclitic for nitrophenylphosphocholine and (iii) fall to bind to or provide protection against S. pneumoniae. Single prototypic V-D variants of the T15 idiotype (Asp95H), M603 idiotype (Asn95H) and M167 idiotype (Asp95H-Ala96H) were also induced in Xid mice. The M603-like and M167-like antibodies bound to and protected against S. pneumoniae even though they exhibited Kas for PC which were lower than T15 idiotype+ antibodies. These data demonstrate that small changes in the V-D junctional sequence of the T15 (VH1) heavy chain alter L chain usage and the structure of the PC binding site so that the PC expressed on S. pneumoniae is no longer recognized.

GM and KM allotypes in eight tribal populations of Madyha Pradesh and Orissa, India.

Serum samples from eight endogamous Indian tribal populations of Madhya Pradesh (Dhurwa, Halba, Bhatra, Muria, Maria) and Orissa (Deshia Khond, Binjhal, Kisan) with a total of n = 731 unrelated individuals were typed for G1M (1,2,3,17), G3M (5,10,11,13,14,15,16,21, 26), and KM (1). In seven of these populations five different GM haplotypes were found: GM* 1,17;21,26; GM* 1,17;10,11,13,15,16; GM* 1,2, 17;21,26; GM* 1,3;5,10,11,13,14,26; and GM* 3;5,10,11,13,14,26. In the Kisan sample the haplotype GM* 1,2,17;21,26 is absent. The intergroup variability in the distribution of these haplotypes is considerable and statistically highly significant. The reasons for that can be attributed to the ethnohistory and to the genetic isolation of these eight endogamous tribal populations. The GM haplotype distribution pattern of all these groups is quite different from that of the non-tribal populations of India, whereas it is in good agreement with that of the so far tested other tribal populations from India. This can be explained by different origin and history of the Indian tribal and non-tribal populations. In the KM system, too, remarkable variability is seen in the distribution of phenotype and allele frequencies among the eight tribal populations under study.

IgE allotypes in sera of mice with autoimmune diseases and in mice with graft-versus-host disease after transfusion or bone marrow transplantation.

Isotype- and allotype-specific IgE determinations by sandwich ELISA with monoclonal antibodies to Igh-7 (6HD5, HMK-12), Igh-7a (No. 297) and Igh-7b /JKS-6) were used to qualitate antibodies of the IgE class. The sensitivity of the method is 0.1 ng/ml. Serum IgE levels were much higher in mice infected with Nippostrongylus brasiliensis. In sera of BALB/c, AKR and C3H/JCL mice only IgEa, in sera of C57BL/6 and CB-20 only IgEb, in sera of (BALB/c x C57BL/6) F1 mice, both allotypes were detected, as expected. In mice with autoimmune diseases, such as NZB, NZW, (NZB x NZW) F1, MRL/Ipr and MRL/n strains which all belong to the IgEa allotype group, IgE and IgEa were very high. In the sera of BXSB mice, both IgEa and IgEb were detected. Both IgEa and IgEb from donor and host were increased in the sera of mice with graft-versus-host disease after transplantation but only the IgEa (from the donor) was increased in mice with graft-versus-host disease after bone marrow transplantation. In the former, both B cells (from donor and host) secreted IgE, whereas in the latter only the donor cells did. These are the first observations showing the importance of IgE allotype secretion as an indication of graf-versus-host disease.

Immunoglobulin allotypes and genetic susceptibility to invasive Haemophilus influenzae type b and Staphylococcus aureus infections in South African children.

OBJECTIVE: The purpose of this study was to determine whether the G2m(n), G1m(f) and Km(3) immunoglobulin allotypes have any association with susceptibility to invasive Haemophilus influenzae type b (Hib) and Staphylococcus aureus (S. aureus) infections in children. METHODS: Direct enzyme-linked immunosorbent assays with commercially available monoclonal antibodies were established to quantitate G2m(n) and G1m(f) allotypes. A qualitative enzyme-linked immunosorbent assay with polyclonal rabbit anti-Km(3) antibody was established for Km(3) determination. RESULTS: The G2m(n) marker occurred in 34.4% of the mixed ancestry population and 2.9% of the Black population. There was a significantly decreased frequency of the G2m(n) allotype in mixed ancestry children with Hib meningitis (8.5%) and Hib osteomyelitis/septic arthritis and a decreased frequency of Km(3) in black and mixed ancestry children with Hib meningitis. The frequency of G2m(n), G1m(f) and Km(3) allotypes in patients with S. aureus osteomyelitis/septic arthritis were not significantly different from normal population frequency. CONCLUSIONS: This study shows a clear association between the absence of the G2m(n) allotype in mixed ancestry children and susceptibility to invasive infections caused by H. influenzae and an association between the absence of Km(3) and Hib meningitis in both black and mixed ancestry children.

The presence of anti-Fc gamma receptor autoantibodies is related to the clinical presentation of primary Sjögren's syndrome.

OBJECTIVE: Fc gamma receptor III (Fc gamma RIII) is one of the 3 structurally distinct families of receptors for the Fc domain of IgG, and its Fc gamma RIIIb isoform is exclusively expressed in polymorphonuclear (PMN) cells. We sought to detect anti-Fc gamma RIII autoantibodies in serum from patients with primary Sjögren's syndrome (SS). METHODS: Sixty-six patients with SS and 44 healthy controls were enrolled in the study. ELISA were developed. RESULTS: IgG and IgM autoantibodies were found in 16 (10 IgG+ IgM+ and 6 IgG+ IgM-) and 24 patients (10 IgG+ IgM+ and 14 IgG- IgM+) respectively. Their frequency was higher in patients with nonerosive arthritis (p < 0.02), Raynaud's phenomenon (p < 0.003), and lung involvement (p < 0.02) than in patients without such complications. The levels of IgM and IgG antibody (p < 0.05) correlated with the content of IgA without the circulating immune complex (IC), while there was no relationship between anti-Fc gamma RIII activity and the PMN count. CONCLUSION: Anti-Fc gamma RIII autoantibodies may act as an acquired additional factor further compromising IC handling in individuals who share HLA-DR3 alloantigen.

Immunoglobulin allotypes (GM and KM) and their interactions with HLA antigens in autoimmune diseases: a review.

GM and KM immunoglobulin (Ig) allotypes and their interactions with HLA antigens have been analyzed in various autoimmune diseases: multiple sclerosis, rheumatoid arthritis, insulin-dependent diabetes mellitus (IDDM), systemic lupus erythematosus, coeliac disease, Crohn's disease, Graves' disease, atrophic thyroiditis, Hashimoto's thyroiditis, myasthenia gravis, chronic active hepatitis, alopecia areata, uveitis, vitiligo, Turner's syndrome, glomerular nephritis, Berger's disease and idiopathic dilated cardiomyopathy. This review reports published results about associations or linkages, as well as the origins of the populations, the numbers of patients and controls tested. The possible role of Ig polymorphisms in the physiopathology of autoimmune diseases is discussed. Ig allotypes and statistical methods used to analyse the HLA and Ig data are also described.

Monoclonal antibodies against the Fc fragment of IgA alpha-chain: preparation and clinical application.

Fragments of the heavy alpha-chain of IgA (IgA Fc alpha) were prepared and purified from the serum of patients with heavy chain disease. It was used to immunize BALB/c mice. The immunized mouse spleen cells were then fused with myeloma cells Sp2/O. Through successive recloning of the hybridoma cell lines by the limiting dilution method, four subclonal hybridoma cells (D2, E4, F5 and G10) capable of screening IgA Fc alpha were found. Intraperitoneal transplantation of these cells induced ascites in the BALB/c mice, from which four types of monoclonal antibodies (McAb-IgA Fc alpha) were obtained. The highest McAb titer was as high as 1:16348 by the hemagglutination inhibition (HAI) test. These McAb's reacted specifically with IgA Fc alpha, but not with IgG, IgM, IgD, kappa or lambda. Serum samples from 92 normal subjects and 73 patients with various diseases were tested for Ig genetic markers by means of HAI. The family trees of three positive cases were surveyed. Results showed that the McAb-IgA Fc alpha may be used to determine the genetic marker of IgA allotypes.

Clonal characterization of the human IgG antibody repertoire to Haemophilus influenzae type b polysaccharide. V. In vivo expression of individual antibody clones is dependent on Ig CH haplotypes and the categories of antigen.

Antibodies (Ab) to the polysaccharide capsule of Haemophilus influenzae type b (Hib-PS) provide protection against Haemophilus influenzae type b disease in children, and Hib-PS vaccines with different immunologic properties are widely used clinically. The repertoire of human anti-Hib-PS Ab induced by these vaccines is relatively restricted and can be divided into two types by the structure of the light chain V region. Ab using A2-V kappa II gene product, which account for the majority of anti-Hib-PS Ab response in most patients, show little somatic mutations. In contrast, non-Ab using A2-V kappa II gene product use VL genes from the V kappa I, V kappa II, V kappa III, V kappa IV, and V lambda subgroups, are variably expressed among patients, and contain somatic mutations. To further study the expression of these two types of anti-Hib-PS Ab, we have produced KB13, a mAb specific for V kappa II subgroup, and used mAb specific for various other VL subgroups to develop immunoassays specific for anti-Hib-PS Ab of each VL subgroup. When Ig allotypes were studied for the effect on the Ab repertoire, A2-V kappa II (A2) Ab were found to be expressed less in patients expressing fb or zag CH haplotypes (p < 0.05). When the T cell-independent Hib-PS carbohydrate vaccine was compared to two T cell-dependent Hib-PS protein conjugate vaccines for their effect on Ab repertoire, Ab using V kappa III VL were found to be more often elicited with the conjugate vaccines than with the Hib-PS carbohydrate vaccine (p < 0.01). Thus, individual members of the anti-Hib-PS Ab repertoire differ not only in their V region structure but also in the control of their expression.

Expression of immunoglobulin kappa and lambda chains in mink.

The ratio of kappa and lambda chains of immunoglobulins varies significantly from one species to another. It has previously been thought that lambda was only type expressed in mink. We tested mink immunoglobulin light chains using two monoclonal antibodies G80 and G88. It has been shown that G80 and G88 specifically recognize two antigenically different subpopulations of the light chains. Immunochemical analysis of these subpopulations separated by affinity chromatography suggested that they represent lambda and kappa types of light chains, respectively. Screening of a mink cDNA library with monoclonal antibody G88 resulted in the isolation of clone pIGK-1 containing kappa chain-encoding sequence. The cDNA insert of pIGK-1 included most of the V segment, as well as the J, C and 3' untranslated sequences. Mink V kappa sequence shown the highest homology with the human V kappa II subgroup genes (76-79%). Mink C kappa sequence was 53-63% homologous to C kappa of other species. The striking feature of mink C kappa chain is the presence of glutamine in the C-terminal position. Southern blot analysis suggested that mink haploid genome has one C kappa gene and multiple V kappa genes. The kappa:lambda chain ratio in the 12 minks studied was, on the average, 46:54. The same ratio was observed for the kappa- and lambda-producing cells in the mesenteric lymph nodes. The five previously identified mink light chain allotypes were assigned to the lambda chains, thereby confirming that lambda chains in this species are additionally subdivided into several subtypes.

HLA antigens and complement C4 allotypes in patients with chronic biologically false positive (CBFP) seroreactions for syphilis: a follow-up study of SLE patients and CBFP reactors.

We report a follow-up of our previous study of HLA markers in 118 unrelated patients: 49 with definite systemic lupus erythematosus (SLE) (group 1), 32 with definite or probable SLE and chronic biologically false positive (CBFP) seroreactions for syphilis (group 2), and 37 CBFP reactors (group 3). Definite SLE was confirmed in 28 (90.3%) of the patients in group 2, equally in HLA B8- and HLA B7-positive patients. Three of the CBFP reactors developed SLE, two (40%) out of five HLA B8-positive as compared to one (6.6%) out of 15 HLA B7-positive CBFP reactors (P = 0.07). Fourteen patients died (groups 1 and 2). Eight of the 24 HLA B8-positive patients died in contrast to one of the 20 HLA B7-positive patients (P < 0.02). Of the CBFP reactors, 70.9% had complement C4 null alleles as compared to 47.9% in controls (P = 0.05) and 50% had C4A null alleles as compared to 17.8% in controls (P < 0.05). C4B null alleles were found in 28.6% (28.6% in controls, P is not significant). The null alleles for C4A were not solely in a linkage disequilibrium with the HLA B8 DR3 haplotype. CBFP reactors with C4A null alleles had a higher risk of developing SLE, lupus-like disease or symptoms such as photosensitivity, cutaneous vasculitis and/or autoantibodies than did those with no C4A null alleles (P < 0.02).

Genetic, immunologic and biotransformation studies of patients on procainamide.

This report represents follow-up observations of a unique long-term study of patients on procainamide (PA) for various cardiac arrhythmias. Serologic and clinical evaluations associated with drug-related autoimmunity were assessed and patients were characterized for factors postulated to influence susceptibility to autoimmunity, including acetylator phenotype, oxidative metabolism of PA, HLA class profile, and production of interleukin-1 (IL-1) and tumor necrosis factor (TNF). Fifty-two percent had IgM and 70% IgG antibodies to total histones; 67% had IgG antibodies to histone H2A/H2B. Patients were equally divided between fast and slow acetylators. N-oxidative metabolism of PA was indicated by the presence of urinary nitroprocainamide, which correlated with elevated titers of antihistone antibodies. There was a significant incidence of the DQw7 split of DQw3 in PA patients when compared to controls, and the frequency of antibodies to total histones and H2A/H2B was significantly increased in the DQw7 patients. C4A*QO and C4B*QO alleles were more frequent in the PA patients than in controls. IL-1 and TNF production was not different in patients compared to controls. These data suggest that certain genetic factors may serve as markers for PA-related autoimmunity.

T cell-induced suppression of IgG2ab expression in vivo leads to a large reduction of C gamma 2ab mRNA levels.

In the T cell-induced suppression of IgG2ab expression, the level at which B cells are blocked in their development to IgG2ab-producing plasma cells was investigated. Although IgG2ab+ lymphocytes were barely detected in normal and IgG2ab-suppressed mice, intracellular IgG2ab was only detected in crude cell extracts from normal mice. B lymphocytes producing IgG2ab were revealed in T cell-depleted splenocytes from normal mice (86 +/- 15/10(6) cells), whereas corresponding cell preparations from IgG2ab-suppressed mice were completely free of such lymphocytes. However, in vitro stimulation of cell preparations from both normal and IgG2ab-suppressed mice with LPS plus rIFN-gamma resulted in IgG2ab production. Accounting for differences in spleen size between the two types of mice, these stimuli induced comparable cell proliferation and numbers of IgG2ab-producing lymphocytes. In addition, the level of IgG2ab production per cell was similar in the two types of stimulated cells. This demonstrates that normal and IgG2ab-suppressed mice have the same potential to generate IgG2ab-producing cells. By using a sensitive and specific RNase protection assay, C gamma 2ab transcripts were detected in total RNA preparations from IgG2ab-suppressed mice. The levels of C gamma 2ab gene expression in spleen were much lower (between 150 and 400 times less) in IgG2ab-suppressed mice than in normal mice. Taken together, our data suggest that B lymphocytes committed to IgG2ab production represent the target of CD8+ T cells, which we have previously shown to be required for suppression. The target B cells are very efficiently and rapidly silenced, as demonstrated by the absence of detectable serum IgG2ab and corresponding low levels of C gamma 2ab mRNA.

Immunoglobulin allotypic markers in Kawasaki disease.

We investigated the possible relationship of the distribution of immunoglobulin allotypic markers for susceptibility to Kawasaki disease in Japanese, Japanese-American, and white American populations. The kappa-chain allotype Km1 was present in 25.6% of sera from white patients with Kawasaki disease and in 14.4% of control sera (p < 0.01), and the combination of Km1 with Gm heterozygosity was present in 17.9% of white patients with Kawasaki disease and in 6.4% of control sera (p < 0.0001). In all populations studied, differences were observed between the patients with Kawasaki disease and the race-matched control subjects. The findings support the hypothesis that one or more unknown infectious agents may trigger genetically influenced immune responses that result in clinically recognizable Kawasaki disease.

MRL/lpr-->severe combined immunodeficiency mouse allografts produce autoantibodies, acute graft-versus-host disease or a wasting syndrome depending on the source of cells.

MRL/lpr (lpr) mice spontaneously develop a lupus-like illness as well as massive lymphadenopathy. Attempts to transfer autoimmunity by adoptive transfer or radiation bone marrow chimeras have been unsuccessful. Since severe combined immunodeficiency (SCID) mice have been engrafted with human and rat xenografts without apparent graft-versus-host disease (GVHD), we subjected SCID mice to low-dose irradiation and reconstituted the mice with spleen cells from young or old lpr mice or with lpr bone marrow. Fourteen out of twenty (70%) of SCID mice engrafted with spleen cells from old lpr mice produced autoantibodies (anti-DNA and anti-Sm) without evidence of the severe lymphoid atrophy previously described for lpr spleen-->+/+ chimeras. SCID mice engrafted with spleen cells from young lpr mice developed acute GVHD and 5/6 (83%) died within 4 weeks post-transfer. Although 8/11 (73%) of lpr-->SCID bone marrow allografts survived for at least 4 months, these mice developed a wasting disease characterized by lymphoid atrophy and fibrosis without the production of autoantibodies. None of the lpr-->SCID grafts resulted in the transfer of double negative T cells or the lymphoproliferative syndrome characteristic of MRL/lpr mice. These findings indicate that SCID mice can be engrafted with splenocytes from old MRL/lpr mice and that B cells continue to secrete autoantibodies for several months in the SCID recipients. This study also demonstrates that, unlike i.p. transplant of xenogeneic cells, acute GVHD is a consistent feature of i.p. transplants of normal allogeneic mononuclear cells into SCID mice.