Training-induced changes in physical performance can be achieved without body mass reduction after eight week of strength and injury prevention oriented programme in volleyball female players.

The purpose of the study was to analyse the changes in muscle strength, power, and somatic parameters in elite volleyball players after a specific pre-season training programme aimed at improving jumping and strength performance and injury prevention. Twelve junior female volleyball players participated in an 8-week training programme. Anthropometric characteristics, isokinetic peak torque (PT) single-joint knee flexion (H) and extension (Q) at 60º/s and 180º/s, counter movement jump (CMJ), squat jump (SJ), and reactive strength index (RSI) were measured before and after intervention. Significant moderate effects were found in flexor concentric PT at 60º/s and at 180 º/s in the dominant leg (DL) (18.3±15.1%, likely; 17.8±11.2%, very likely) and in extensor concentric PT at 180º/s (7.4%±7.8%, very likely) in the DL. In the non-dominant leg (NL) significant moderate effects were found in flexor concentric PT at 60º/s and at 180º/s (13.7±11.3%, likely; 13.4±8.0%, very likely) and in extensor concentric PT at 180º/s (10.7±11.5%, very likely). Small to moderate changes were observed for H/QCONV in the DL at 60º/s and 180º/s (15.9±14.1%; 9.6±10.4%, both likely) and in the NL at 60º/s (moderate change, 9.6±11.8%, likely), and small to moderate decreases were detected for H/QFUNC at 180º/s, in both the DL and NL (-7.0±8.3%, likely; -9.5±10.0%, likely). Training-induced changes in jumping performance were trivial (for RSI) to small (for CMJ and SJ). The applied pre-season training programme induced a number of positive changes in physical performance and risk of injury, despite a lack of changes in body mass and composition.

Are genes encoding proteoglycans really associated with the risk of anterior cruciate ligament rupture?

Proteoglycans are considered integral structural components of tendon and ligament and have been implicated in the resistance of compressive forces, collagen fibrillogenesis, matrix remodelling and cell signalling. Several sequence variants within genes encoding proteoglycans were recently implicated in modulating anterior cruciate ligament ruptures (ACLR). This study aimed to test the previously implicated variants in proteoglycan and vascular epithelial growth factor encoding genes with risk of ACLR in a population from Poland. A case control genetic association study was conducted using DNA samples from 143 healthy participants without a history of ACL injuries (99 male and 44 females) (CON group) and 229 surgically diagnosed ACLR participants (158 males and 71 females). All samples were genotyped for the ACAN: rs1516797, BGN: rs1042103, rs1126499, DCN: rs516115 and VEGFA: rs699947 variants. Main findings included the (i) ACAN rs1516797 G/T genotype which was underrepresented in the CON group (CON: 36%, n=52, ACLR: 49%, n=112, p=0.017, OR=1.68, 95% CI 1.09 to 2.57) when all participants were investigated and (ii) the BGN rs1042103 A allele was significantly under-represented in the male CON group compared to the male ACLR group (CON: 39%, n=78, ACLR: 49%, n=156, p=0.029, OR=1.5, 95% CI 1.05 to 2.15). Furthermore, BGN inferred haplotypes were highlighted with altered ACLR susceptibility. Although the study implicated the ACAN and BGN genes (combination of genotype, allele and haplotype) in modulating ACLR susceptibility, several differences were noted with previous published findings.

16(th) IHIW: report of the MICA project.

The MICA Project of the 16th International HLA and Immunogenetics Workshop was planned to improve the detection of specific antibodies against MICA antigens that develop in organ transplant recipients. It was organized as a serum exchange with 18 laboratories located in seven countries as participants. Each laboratory used the MICA screening reagents available to them. It was found that there were four different kits of Luminex beads conjugated with MICA antigens that could be used in these experiments. They were, kit BL, produced by Rainer Blasczyk in Hannover, Germany, containing 12 MICA antigens, the kit from Gen-Probe (GP), which included 28 MICA antigens, the beads from One Lambda (OL), consisting of ten antigens, and the beads produced in the laboratory of the organizers (ZS) consisting of 11 MICA antigens. The sera were all from transplant recipients and represented a spectrum of MICA-specific antibodies that are commonly found. All laboratories accurately could recognize which sera contained MICA antibodies. None failed to recognize the negative serum that was included in one of the shipments. While there were important differences in the specificities of MICA antigens recognized by the different kits, the results in laboratories using the same beads were remarkably similar. Feedback was given to the participants, and especially to the producers of the antibody detection kits, after each set of four sera was sent, and results were collected. Certain beads were replaced and results improved as can be seen in the results of the third shipment of sera. It is important for laboratories to be able to accurately determine the specificity of antibodies against MICA to know whether the antibodies are donor specific. Although complete consensus was not attained, some important errors were corrected, and a better understanding of how to accurately determine MICA allele-specific antibodies was developed.

Antagonist activation exercises elicit similar post-activation performance enhancement as agonist activities on throwing performance.

BACKGROUND: This study aimed to determine the acute effect of agonist and antagonist conditioning activities (CA) on medicine ball throw performance among female softball players. METHODS: Thirteen national-level female softball players (age 22.2 ± 3.1 years; body mass 68.3 ± 11.3 kg; softball experience 7.3 ± 2.4 years) performed 3 medicine ball chest throws before conditioning activity (CA) and after CA respectively in 3rd, 6th, and 9th minute. CA was the bench press and bent-over barbell row with 2 sets of 4 repetitions at 60% and 80% of one-repetition maximum, and 2 sets of 4 repetition bodyweight push up. RESULTS: Two-way ANOVA revealed an increase in throwing distance (p < 0.001) after bent over barbell row and push-up exercise, and an increase in throwing speed (p < 0.001) after bench press and push-up. All performance increases were in moderate effect size (Cohen d 0.33-0.41), and no differences were found between the experimental CA. CONCLUSIONS: We conclude that upper body throwing performance is similar after antagonist exercise and agonist CA, both agonist and antagonist CA increase muscle power. In the resistance training practice, we recommend the interchange of agonist and antagonist CA using bodyweight push-up or submaximal intensity (80% of 1RM) bench press and bent over barbell row to succeed post-activation performance enhancement in upper limbs.

HLA-D region-associated determinants serve as targets for human cell-mediated lysis.

Effector cells for cell-mediated lysis (CML) were generated by in vitro culture of lymphocytes from selected donors with X-irradiated cells from unrelated subjects who were HLA-D homozygous and matched to the responders for the antigens of the HLA-A and HLA-B regions. By using chromium labeled monocytes as target cells, cytotoxicity was found to correlate with presence of HLA-D region antigens matching those of the stimulating cells. Such CML reactions apparently directed at products of HLA-D, were inhibited by addition of unlabeled monocytes or B lymphocytes. These unlabeled cells had to be matched for HLA-D with the stimulating cells used to generate the effector populations. The results suggested that products of HLA-D, perhaps the DR antigens, were recognized by cytotoxic lymphocytes.

Restriction fragment length polymorphism of the gene encoding the alpha chain of the human T cell receptor.

Two allelic forms of the T cell antigen receptor alpha chain gene were discerned by restriction fragment length polymorphism (RFLP) employing the T cell antigen receptor alpha chain probe pGA5, and the restriction enzyme Bgl II. Analysis revealed that the polymorphic fragments are detected by a probe specific for the constant region exon of the T cell antigen receptor alpha chain gene. Furthermore, the polymorphic fragments were shown to segregate within families. The two allelic forms yield two homozygous states, 3.2/3.2 and 2.9/2.9, at a frequency of 76.5 and 2.9%, respectively, within the normal population. The heterozygous state was observed in 20.6% of the population. The discovery of allelic forms of both the alpha and beta chains of the T cell antigen receptor genes may provide a unique opportunity to study heritable markers of T cell function in several human diseases.

Molecular localization of human class II MT2 and MT3 determinants.

The specificities of the monoclonal antibodies I-LR2 and 109d6, which recognize MT2- and MT3-like serologic determinants, respectively, have been confirmed by panel testing. In addition, the relationships of these antibodies to other monoclonal antibodies and alloantisera have been studied by means of cell surface fluorescence, complement-dependent cytotoxicity and immunoprecipitation. Using these monoclonal antibodies, molecules encoded by the HLA-D region have been isolated and characterized by amino acid sequencing and peptide mapping. By these criteria, the major populations of molecules bearing MT2- and MT3-like determinants are indistinguishable from DR molecules.

Structural analysis of a human I-A homologue using a monoclonal antibody that recognizes an MB3-like specificity.

Monoclonal antibody IVD12 was used to isolate and characterize a human Ia molecule present on B cells that generally display DR4 or DR5 phenotypes. The specificity of binding of IVD12 to human peripheral blood B cells from 75 normal individuals and 19 homozygous human lymphoblastoid B cell lines was identical to the supertypic specificity MB3 previously defined. Furthermore, IVD12-reactivity was shown to segregate with HLA in three informative families. In each family, individuals positive for IVD12 binding were also positive for DR4 or DR5. Using IVD12, a molecule has been isolated from the homozygous cell line PRIESS (DR4/4) and has been shown by amino acid sequence analysis to be homologous to the murine I-A and human HLA-DS molecules. These findings suggest that the MB3 specificity is found on a molecule encoded by loci distinct from those loci which encode HLA-DR molecules. This molecule represents the third family of HLA-D region molecules isolated from the cell line PRIESS. Both HLA-DR and HLA-SB molecules from this cell line were previously shown by amino acid sequence analysis to be I-E-like but distinct from one another. Collectively, these data provide evidence that the HLA-D region contains at least six loci encoding distinct alpha and beta chains for the HLA-SB, HLA-DR, and HLA-DS molecules.

Role of MICA in the immune response to transplants.

Among the cell surface antigens that can elicit an immune response in transplant recipients MICA antigens occupy a special place. They are similar to human leukocyte antigens (HLAs) while being very different from them. They are not as polymorphic and their quantity is smaller. In consequence, the impact of MICA antigens in transplantation is not as dramatic. However, our early guess that these ligands of NKG2D could elicit antibodies and cell-mediated immunity has been definitely confirmed. Careful analysis with MICA transfectant cells, for absorption and elution, established the specificity of the epitopes involved. Typing of recipients and donors by sequencing the MICA alleles has established that de novo antibodies produced in kidney transplant recipients are directed at mismatched MICA epitopes and are associated with acute rejection and chronic transplant failure. Acute graft-versus-host disease was observed in stem cell recipients who were mismatched for MICA.

HLA class I antibodies provoke graft arteriosclerosis in human arteries transplanted into SCID/beige mice.

Antibodies toward HLA class I and/or MICA are commonly observed in transplanted patients suffering from allograft arteriosclerosis, also called chronic vascular rejection (CVR). The relative importance of cellular versus humoral alloreactivity for CVR is still disputed. We demonstrate that antibodies toward HLA class I provoke lesions typical for CVR in human arteries in vivo in the absence of cellular immunity. To show this, we grafted segments of human mesenteric arteries from 8 deceased organ donors into 36 immunodeficient SCID/beige mice in the infrarenal aortic position. Three mice died postoperatively. The remaining 33 mice received weekly i.v. injections of either a monoclonal antibody toward HLA class I, toward MICA or an irrelevant monoclonal antibody. At sacrifice after 6 weeks, mice receiving the HLA antibody showed a significant neointimal thickening in the grafted artery due to smooth muscle cell (SMC) proliferation while control mice receiving anti-MICA or irrelevant antibody showed little or no thickening. Whereas antibodies toward HLA class I were mitogenic to SMC in vitro, those directed toward MICA did not have any effect. Humoral alloreactivity toward HLA may thus play a causal role for the development of CVR and this opens new possibilities for the treatment of CVR.

HL-A antigens in mummified pre-Columbian tissues.

Tissue extracts of pre-Columbian mummies, from 500 to 2000 years old, were found to inhibit specific antibodies to HL-A. Two-thirds of the specimens tested gave positive results. Patterns of reactions obtained with different antiserums detecting the same antigen were concordant and consistent with known relations between HL-A antigens. The distribution of antigens found was similar to that observed in present-day descendants of the ancient populations studied. Although artifacts due to contaminating substances could have occurred, the reactions resembled in many respects those of HL-A antigens rather than those of nonspecific cross-reacting inhibitors. Development of a technique for HL-A typing of mummified remains may open new possibilities for anthropologic studies.

Gene for familial psoriasis susceptibility mapped to the distal end of human chromosome 17q.

A gene involved in psoriasis susceptibility was localized to the distal region of human chromosome 17q as a result of a genome-wide linkage analysis with polymorphic microsatellites and eight multiply affected psoriasis kindreds. In the family which showed the strongest evidence for linkage, the recombination fraction between a psoriasis susceptibility locus and D17S784 was 0.04 with a maximum two-point lod score of 5.33. There was also evidence for genetic heterogeneity and although none of the linked families showed any association with HLA-Cw6, two unlinked families showed weak levels of association. This study demonstrates that in some families, psoriasis susceptibility is due to variation at a single major genetic locus other than the human lymphocyte antigen locus.

Mixed lymphocyte cultures in rheumatoid arthritis.

Random one-way microtiter mixed lymphocyte cultures between 43 rheumatoid arthritis (RA) patients and 45 controls consisting of 26 normal subjects and 19 miscellaneous non-RA patients were performed and results were evaluated as relative responses. Low responses (consisting of relative response less than 38%) were found in 31 out of 43 RA patients in cultures against eight of the RA stimulators. The remaining 35 RA stimulators tested yielded only normal mixed lymphocyte culture reactions. The same RA patients used as responders never produced low responses when stimulated by non-RA lymphocytes. But six of the control subjects gave low responses to two RA stimulators. The low responses did not appear to correlate with intake of aspirin, prednisolone, or gold salts, nor could they be reproduced by addition of RA serum of 7S or 19S fractions thereof containing either polyclonal or monoclonal rheumatoid factors. Short-term culture and washing before mixing with the allogeneic cells did not change the low responses suggesting that in vivo bound autoantibodies against lymphocyte receptors were not involved. Study of the inheritance of HLA and mixed lymphocyte culture determinants in the family of patient A. C. who most frequently elicited low responses indicated she was homozygous for a lymphocyte-defined determinant which has been called R. The low responses to A. C. could be interpreted as typing responses based on sharing of the same or of a similar lymphocyte-defined determinant. This gene appears to be increased in patients with RA with respect to non-RA controls and may reflect an association of genes within the HLA chromosomal region leading to predisposition for the development of RA.

Different HLA-D associations in adult and juvenile rheumatoid arthritis.

HLA-D typing was performed in 126 patients with juvenile rheumatoid arthritis. HLA-DW4, the antigen found in previous studies to characterize adult rheumatoid arthritis, had a significantly lower frequency in children with arthritis than in normal controls (P less than 0.04). By contrast, in children the antigens HLA-DW7 (P less than 0.03) and HLA-DW8 (P less than 0.01) were increased compared to controls. The antigen TMo, detected with homozygous typing cells from a child with juvenile rheumatoid arthritis, was found to be related to the cross-reactive specificities HLA-DW7 and DW11. 46% of the patients with persistent pauciarticular arthritis of childhood typed for the antigen TMo, compared to only 1% of normal controls. Thus the relative risk for persistent pauciarticular arthritis in relation to the presence of TMo was 67.7 (P less than 0.0001). These results provide evidence of fundamental differences between adult rheumatoid arthritis and arthritis of childhood. The latter group appears to include a population distinguishable clinically and characterized in these studies by the HLA-D determinant TMo.

A new antigen system expressed in human endothelial cells.

Kidney transplant recipients were previously found to have antibodies that reacted with cells isolated from the endothelium of umbilical cord veins and which were not cytotoxic for lymphocytes from the same donors. Results of the present experiments indicate that endothelial (E) antigens are different from previously known HLA antigens and also from Ia-like antigens of bone marrow-derived (B) lymphocytes. Attempts to absorb E antibodies with lymphocytes from E-positive donors failed in most cases. Antigen redistribution experiments showed that E antigens were located in separate molecules from the products of HLA-A, B, and C. Thus, E cells treated with E antibody became resistant to lysis by the antibody used, but remained susceptible to the effects of typing sera for alleles of HLA-A, B, and C. Antibodies to E cells were also cytotoxic for blood monocytes. Moreover, monocytes were able to absorb E-antibody reactions, indicating that similar antigens were expressed in both cells. E antibodies did not react with B lymphocytes isolated from peripheral blood. In that regard E antibodies were different from antibodies to human Ia-like antigens which reacted with E cells, monocytes, and isolated B lymphocytes. Thus it appears that E antigens constitute a system of human alloantigens which has not been previously identified. The possibility that these antigens play a role in kidney allograft rejection should now be investigated since matching can be performed using monocytes isolated from the blood of recipients and donors.

Human histocompatibility antigen associations in subacute cutaneous lupus erythematosus.

We have identified a clinically distinct subset of lupus erythematosus patients marked by the presence of a histologically proven, nonscarring variety of cutaneous LE (subacute cutaneous LE) in which there is a very high frequency of the human leukocyte antigens (HLA) B8 and DR3. Differences in the configuration of their skin lesions allowed a separation of the patients into two clinical subgroups; annular and papulosquamous. HLA-B8 was increased in the annular subgroup (81%, corrected P (Pc) < 0.007) and combined group (65%, Pc < 0.004). HLA-DR3 was present in all 11 of the annular patients (10%, Pc < 0.00008). In addition, HLA-DR3 was present in increased frequencies in the papulosquamous subgroup (60%, Pc < 0.04) and combined group (77%, Pc < 0.00008). Thus, HLA-DR3 positive individuals have a relative risk of 10.8 for developing subacute cutaneous LE of either type and an even greater relative risk (67.1) for the annular variety. The HLA phenotype A1, B8, DR3 was also found more commonly in the annular (73%, P < 0.00008) and combined patient groups (46%, P < 0.004). These HLA associations, which are stronger than ever before reported for any form of LE, did not result from the concurrent presence of subclinical Sjögren's syndrome. Thus, subacute cutaneous LE can now be added to the growing list of HLA-B8, DR3-associated diseases that have autoimmune features.

Specificities of antibodies eluted from human cadaveric renal allografts. Multiple mechanisms of renal allograft injury.

The purpose of the present experiments was to evaluate the role of circulating antibodies in the rejection of human renal allografts and to study the apparent target(s) for antibody binding. Eluates obtained from surgical biopsy and nephrectomy specimens of rejecting, cadaveric human renal allografts were tested for antibodies directed to structural antigens of normal kidney and for cytotoxic antibody activity against mononuclear cell populations. By indirect immunofluorescence 23 of 35 eluates contained immunoglobulin that bound to normal kidney. Staining was in smooth muscle only in 10 patients, in smooth muscle and other structures such as tubular basement membranes, proximal cells, or brush border in 9 patients, and in structures other than smooth muscle in 4 patients. All 16 eluates tested contained antibodies cytotoxic for cells derived from a panel of normal volunteers. Six were cytotoxic to T cells and 10 to B cell and monocyte-enriched preparations. Absorption of eluates with pooled buffy coat cells, platelet concentrates and packed, cultured B cells removed antibodies reactive with vascular wall smooth muscle and endothelium, but not antibodies to tubular basement membranes, proximal or distal tubular cells, brush border, or other structures of kidney sections. Two of five eluates containing antikidney antibodies were found to bind to rat kidneys in vivo. These results suggest that circulating antibodies participate in cadaveric renal allograft destruction and demonstrate that they can be recovered directly from the allograft. Moreover, the data indicate that there are different antibody populations involved: some clearly directed to allo-specific differences and others that are apparently kidney-specific.

High frequency of histocompatibility antigens HLA-DR3 and DR4 in herpes gestations.

Herpes gestationis (HG) is a rare, autoimmune, vesiculobullous disease of pregnancy or the puerperium characterized by the deposition of complement (and occasionally immunoglobulin) within the lamina lucida of the cutaneous basement membrane zone. We have studied 23 patients with a history of HG, 20 of whom had typical immunofluorescence findings during the active phase of their disease. HLA typing showed HLA-DR3 in 61% of patients (controls 22%, Pc less than 0.005) and the combination of DR3, DR4 in 43% (controls 3%, Pc less than 0.00001). The most striking finding of this study was that the greatest risk of HG is associated with the concurrent presence of two specific histocompatibility leukocyte antigen (HLA)-DR antigens.

Immunoglobulin in clinically uninvolved skin in systemic lupus erythematosus: association with renal disease.

23 of 42, or 55%, of patients with systemic lupus erythematous had immunoglobulin deposits along the epidermal basement membrane of uninvolved skin (positive lupus band test [LBT]). In patients with low serum complement levels, 91% had a positive LBT), as compared with 15% in those with normal complement levels. The LBT was positive in 70% of patients with clinical and laboratory evidence of renal disease, but in only 31% of patients without renal disease. 81% of patients with the more severe histologic forms of lupus nephritis, i.e., proliferative glomerulonephritis and membranous glomerulonephritis, and positive tests, whereas only 23% with mesangial glomerulitis or normal histologic findings were positive. Immunoglobulins of the same class found in the skin were detected in the glomeruli of patients examined by renal biopsy. These results suggest that there is a relationship between the occurrence of immunoglobulin in the epidermal basement membrane and the presence of the more severe forms of lupus nephritis.

Development and characterization of desmoglein-3 specific T cells from patients with pemphigus vulgaris.

Pemphigus vulgaris (PV) is a cutaneous autoimmune disease characterized by blister formation in the suprabasilar layers of skin and mucosae and anti-desmoglein-3 (Dsg3) autoantibodies bound to the surface of lesional keratinocytes and circulating in the serum of patients. This disease can be reproduced in neonatal mice by passive transfer of patients' IgG, indicating that humoral immunity plays an important role in the pathogenesis of PV. Currently, the role of T lymphocytes in the development of PV is not clear. Here, we report that three immunoreactive segments of the ectodomain of Dsg3 specifically induced proliferation of T cells from PV patients. We found that T lymphocytes from 13 out of 14 patients responded to at least one of three Dsg3 peptides. T cells from controls and other patient groups did not respond to these Dsg3 peptides. The major T cell population stimulated by these Dsg3 peptides was CD4 positive. Dsg3-specific T cell lines and clones were developed and were shown to express a CD4 positive memory T cell phenotype. Upon stimulation, these cell lines and clones secreted a Th2-like cytokine profile. The Dsg3 responses of these T cells were restricted to HLA-DR, and not -DQ and -DP, of the major histocompatibility complex. This information will help to elucidate the cellular immune abnormalities leading to production of pathogenic IgG autoantibodies in patients with PV.