bluuebunny/crossref_metadata_2025 huggingface dataset:
IgM anti-idiotypes that block anti-HLA antibodies: naturally occurring or immune antibodies?
Authors: URLACHER A , TONGIO M M , PASQUALI J L
Published: July 2010

Using dithiothreitol (DTT) technique, IgM anti-HLA anti-idiotypic antibodies were detected in a multiparous multitransfused woman. These antibodies were able to inhibit the binding of specific IgG anti-HLA antibodies on their corresponding antigen. The recognized determinants were crossrcactive determinants since they were partially found on anti-HLA antibodies from unrelated individuals. By studying the patient's sera over a period of 2 years, no IgM-IgG switch was observed bui the presence of these antibodies was stable in time, despite the disappearance of the idiotypes (anti-HLA antibodies). However, when looking at the patient's earlier serum, it was shown that these IgM anti-idiotypic antibodies were absent from the first available serum. Thus, these anti-idiotypic antibodies seem to behave both like natural and immune antibodies. The incidence of such antibodies in pretransplant patients is discussed.
Human rib bone marrow mononuclear cells spontaneously synthesize and secrete IgEin vitro
Authors: MACDERMOTT R P , JENDRLSAK G A S , NASH G S , SCHREIBER S , BERTOVICH M J , NAHM M , NONAKA M , FITZGERALD P , KATZ D H , MARCELLETTI J F
Published: July 2010

We have examined spontaneous secretion of IgE by human rib bone marrow mononuclear cells (MNC). Bone marrow MNC from nine out of 12 rib specimens synthesized and secreted substantial amounts of IgE during 14 days of in vitro culture. The 14-day supernatants from these bone marrow MNC contained a mean of 2589 pg/ml of IgR (n= 12) with a maximum production of 1540K pg/ml of IgE compared with small amounts of IgE 80–200 pg/ml) produced by similarly cultured normal and inflammatory bowel disease intestinal lamina propria MNC. Using two rib specimens, time-course studies revealed spontaneous secretion of IgE to be minimal during the first 2 days of culture (152 pg/ml), followed by a steady increase between days 4 (517 pg/ml) and 14 (3588 pg/ml). The addition of pokeweed mitogen resulted in 72% suppression of spontaneous IgE production by bone marrow MNC. The bone marrow MNC isolated from the ribs consisted of 22% Leu 12+ (B) cells of which 3.2% were surface IgE positive. Staining for cytoplasmic immunoglobulin revealed 1% of the bone marrow MNC to be cytoplasmic IgE+. The presence of IgE-bearing and IgE-secreting MNC in human bone marrow is consistent with the observation that allergen-specific IgE-mediated hypersensitivity is adoptively transferred by human bone marrow transplantation and demonstrates the usefulness of human bone marrow MNC for examination of IgE secretory and regulatory events.
Differential effects of CD45 CD45R and CD45R0 monoclonal antibodies in modulating human B cell activation
Authors: DEANE D L , HARVEY E , STEEL C M
Published: July 2010

We have examined the effect of monoclonal antibodies (MoAbs) to different epitopes of the leucocyte common antigen (LCA). CD45, on anti-human immunoglobutin-primed B cell activation. Binding of MoAbs to restricted epitopes present on CD45 glycoproteins of 180 kD and 220 kD (designated CD45R0 and CD45R. respectively) was found to promote B cell proliferation in the presence of T cells. CD45 MoAbs reactive with ‘public’ determinants on all four constituent members of the LCA family (180, 190, 205, and 220 k D) had either little effect or inhibited the basal B cell response to anti-immunoglobulin priming. Simultaneous immunonuorcsccnt analysts of 5-broinodeoxyuridinc incorporation and the expression of CD19 (B cell specific) or CD2 (T cell specific) identified the majority of responder cells as B lymphocytes. CD45R MoAbs significantly enhanced the B cell response to sub-optimal concentrations of interleukin-2. CD45 and CD45R0 MoAbs failed to elicit a similar response. Antibody to the interleukin-2 receptor (anti-Tac) partially blocked the CD45R-driven. T cell-dependent B cell proliferation.
The complement-inhibiting protein, Protectin (CD59 antigen), is present and functionally active on glomerular epithelial cells
Authors: ROONEY I A , DAVIES A , GRIFFITHS D , WILLIAMS J D , DAVIES M , MERI S , LACHMANN P J , MORGAN B P
Published: July 2010

Protectin (CDS9 antigen) is a 20-kD phosphatidyl-inositol-linked membrane protein that inhibits formation of the membrane attack complex (MAC) of complement on homologous cells. Although the antigen has been identified in a number of human tissues, until recently a functional role had been demonstrated only in circulating cells. Using immunofluorescence techniques we have shown the presence of protectin on human glomerular epithelial cells (GEC) in culture and on GEC, tubular epithelial cells and endothelial cells in frozen sections of normal human renal cortex. In addition, we present evidence that this protein functions in protection of GEC from homologous complement: cultured cells incubated with the Fab2 fragment of a monoclonal anti-protectin antibody were markedly more susceptible to killing by homologous serum than were cells in the absence of Fab2 anti-protectin. These findings suggest that this protein may be important in the maintenance of glomerular integrity in vivo, and may be of relevance in certain renal diseases.
Characterization of three monoclonal antibodies to membrane co-factor protein (MCP) of the complement system and quantification of MCP by radioassay
Authors: CHO S-W , OGLESBY T J , HSI B-L , ADAMS E M , ATKINSON J P
Published: July 2010

MCP is a widely distributed regulatory glycoprotein of the complement system which binds C3b and C4b and has factor I-dependent co-factor activity. Monoclonal antibodies raised to lymphocytes (E4.3), chorionic microvilli (GB24) and an embryonal carcinoma cell line (TRA-2–10) recognize MCP (CD46). GB24 inhibited both the binding of MCP to its ligand iC3 and co-factor activity; E4.3 and TRA-2–10 did not. The binding of GB24 to cells bearing MCP was not cross-inhibited by E4.3 or TRA-2–10, but TRA-2–10 blocked binding and displaced pre-bound E4.3. Using these antibodies, we developed a radioassay for quantifying the number of MCP molecules/cell. Human peripheral blood mononuclear (PBMC) and polymorphonuclear cells (PMN) had about 10000 MCP/cell; platelets had about 600/cell, and no MCP was found on erythrocytes. Neoplastic hematopoietic cell lines, of myelocytic and T lymphocytic origin, had several-fold more (20–60000) molecules/cell than peripheral blood cells or B cell lines (about 12000). Malignant epithelial cell lines, HeLa (about 100000/cell) and HEp-2 (about 250000/cell) had the highest MCP expression of any cells examined. These monoclonal antibodies—especially GB24, which blocks MCP function–and the direct binding assay will facilitate the further analysis of the biology of this complement regulatory protein.
Autoantibodies to the URNP particles: relationship to clinical diagnosis and nephritis
Authors: REICHLIN M , VAN VENROOIJ W J
Published: July 2010

Precipitating autoantibodies to the URNP particles were used to select 80 patients, and were further characterized by immunoblotting and quantitative ELISA. These immunochemical results have been related to clinical diagnosis, the frequency of nephritis, and Raynaud's phenomenon. Autoantibodies to the 70-kD polypeptide of the Ul RNP particle were present in 16 out of 19 patients with mixed connective tissue disease (MCTD) and in 27 out of 61 patients with systemic lupus erythematosus (SLE). The ratio of anti-U1 RNP/Sm by ELISA and the frequency of antibody to 70-kD protein were directly related to the frequency of Raynaud's phenomenon and inversely related to the frequency of nehritis.
Increased CDS-positive B lymphocytes in type I diabetes
Authors: MUÑOZ A , GALLART T , VIÑAS O , GOMIS R
Published: July 2010

CD5+ B lymphocytes have been implicated in the production of polyspecific and monospecific antibodies that bind self-antigens, and increased proportions of this B cell subset occur in patients with some autoimmune diseases. We investigated the proportion of peripheral blood CD5+ B lymphocytes in type I diabetic patients. Compared with 18 age-matched healthy subjects. 11 out of 28 (39.2%) type I diabetic patients had increased proportions of circulating CD5+ B lymphocytes with no alterations in the numbers of circulating B and T lymphocytes. Although all patients with increased CD5 B lymphocytes also had serum islet cell antibodies and/or insulin autoantibodies. the occurrence of increased proportions of CD5+ B lymphocytes and scrum autoantibodies was not significantly correlated. Increased proportions of CD5+ B lymphocytes was not related to the time elapsed since the clinical onset of diabetes. In addition, regardless of being increased or normal, the proportion of CD5+ B lymphocytes appeared as a relatively constant phenotype after 1 year of follow-up studies at 3-month intervals in eight patients. Although the significance of these findings remains to be established, the possibility exists that CD5+ B cells play a role in the pathogenesis of type I diabetes.
Induction of tumour necrosis factor-alpha during haemodialysis. Influence of the membrane type
Authors: CHOLLET-MARTIN S , STAMATAKIS G , BAILLY S , MERY J P , GOUGEROT-POCIDALO M A
Published: July 2010

Some of the secondary clinical effects induced by long-term haemodialysis in patients with end-stage renal failure have been related to an increased production of interleukin-1 (IL-1). We investigated the role of another cytokine which shares a number of biological properties with IL-1, tumour necrosis factor-alpha (TNF-α). In long-term haemodialysed patients, we found at the beginning of the dialysis increased plasma TNF-α levels and enhanced monocyte capacity to produce TNF-α spontaneously ex vivo. Non-haemodialysed uraemic patients also presented increased plasma TNF-α levels. During dialysis with cellulose acetate (CA) or polysulphone (PS) membranes, plasma TNF-α levels and the spontaneous and lipopolysaccharide-induced production of TNF-α by monocytes remained at predialysis levels. In contrast, when cuprophane membranes were used, there was a significant increase in plasma TNF-α levels and in both spontaneous (10-fold) and lipopolysaccharide-induced (seven-fold) ex vivo TNF-α production by monocytes. These results suggest that monocytes are stimulated during haemodialysis with the poorly biocompatible cuprophane membrane.
Programmed cell death (apoptosis) in lymphoid and myeloid cell lines during zinc deficiency
Authors: MARTIN S J , MAZDAI G , STRAIN J J , COTTER T G , HANNIGAN B M
Published: July 2010

Three human cell lines of lymphoid (Molt-3 and Raji) or myeloid (HL-60) origin were maintained in vitro under zinc-sufficient or zinc-deficient conditions. Under these conditions, cell proliferation, viability and mode of death (apoptotic or necrotic) were assessed. All three cell types decreased their proliferative capacity and viability under conditions of zinc deficiency. Cell death in the HL-60 and Raji cultures occurred primarily via apoptosis, while most cells in zinc-deficient Molt-3 cultures died via necrosis. Apoptosis in zinc-deficient cultures of HL-60 and Raji cells was characterized by a slow decline in culture viability as cells with condensed and fragmented nuclear DNA appeared. These morphological changes were accompanied by an increase in cell buoyant density, which allowed separation of viable apoptotic cells from their non-apoptotic counterparts by means of percoll step-density gradients. Necrosis in zinc-deficient Molt-3 cultures was characterized by rapid loss of cell culture viability as these cells underwent direct lysis. Intact necrotic cells were easily identified by the flocculated state of their chromatin as well as the decreased basophilia of their cytoplasm. Analysis of DNA from apoptotic HL-60 and Raji cells revealed that internucleosomal DNA degradation, indicative of endogenous endonuclease activation, had occurred, whereas the nuclear DNA of necrotic Molt-3 cells remained relatively unfragmented. The different modes of cell death evoked may reflect the relative sensitivities of cells of these lineages to zinc levels in vivo.
Lack of T cell oligoclonality in enzyme-digested synovial tissue and in synovial fluid in most patients with rheumatoid arthritis
Authors: VAN LAAR J M , MILTENBURG A M M , VERDONK M J A , DAHA M R , DE VRIES R R P , VAN DEN ELSEN P J , BREEDVELD F C
Published: July 2010

The dominant presence or specific T-cell populations in the rheumatoid joint as detected by Southern blot analysis of T cell receptor (TCR) gene rearrangements would indicate local antigen recognition and T cell proliferation. We therefore studied TCR β chain gene rearrangements using a Cβ2 probe in paired samples of T cell populations from synovial tissue and peripheral blood (n= 6) as well as synovial fluid (n= 16) and peripheral blood (n = 18) of patients with rheumatoid arthritis (RA). Peripheral blood mononuclear cells from healthy donors (n= 7) served as a control. T cells were studied directly after isolation or after non-specific expansion with OKT3 monoclonal antibody (MoAb) and T cell growth factor (TCGF). DNA samples were digested with EcoRI and Hind III to detect rearrangements to Cβ1 and Cβ2, respectively. Extra bands were detected in all EcoRI-digested DNA samples prepared from both freshly isolated and non-specifically expanded T cell populations of patients and healthy donors, possibly representing ‘common’ (V-) D-J rearrangements. Dominant rearrangements were found in only two out of 16 synovial fluid T cell populations (one freshly isolated and one expanded) and not in peripheral blood or synovial tissue derived T cell populations. No extra bands were detected in HindIII-digested DNA samples. To investigate the effect of in vitro culture techniques on rearrangement patterns we studied DNA samples prepared from synovial tissue T cells obtained both by outgrowth from tissue with TCGF or by enzyme digestion and subsequent expansion either with TCGF or with OKT3 MoAb and TCGF. Whereas the latter T cell population yielded ‘common’ rearrangements, the former T cell populations yielded different dominant rearrangements. These data indicate that oligoclonality of the T cell populations in synovial tissue and synovial fluid of patients with RA is a rare event. The data also show the influence of in vitro culture techniques on the result of TCR gene rearrangement analysis.
La/SSB ribonucleoprotein levels increased in transformed cells
Authors: ROTHER R P , THOMAS P S
Published: July 2010

Autoantibodies to the La/SSB ribonucleoprotein are commonly found in patients with Sjögren's syndrome. Previous studies have shown that La/SSB accumulates in cells shortly after viral infection. We have extended these studies by investigating levels of the La/SSB antigen in virally and spontaneously transformed cell lines (contact-insensitive and tumourigenic) relative to their non-transformed counterpart cell lines (contact-sensitive and non-tumourigenic). Transformed BALB/ 3T12-3 and KNRK fibroblasts were visibly brighter by immunofluorcscence assay than non-transformed BALB/3T3 and NRK fibroblasts respectively, when reacted with anti-La/SSB specific sera. This was confirmed by flow cytometry, as La/SSB levels were elevated in the transformed counterparts of the same cell lines. An anti-Sm monoclonal antibody and normal human serum reacted with these cell lines failed to show a significant increase by flow cytometry. Finally, a two-fold increase in the La/SSB antigen was demonstrated in cell lysates of these cell lines by a capture ELISA. These data show that La/SSB is elevated in transformed cell lines compared with non-transformed counterpart cell lines and suggest that this increase is not restricted to viral transformation.
Induction of type II collagen-specific antibody production in blood lymphocyte cultures of Rhesus monkeys (Macaca mulatto) with collagen-induced arthritis using the immobilized native antigen
Authors: HART B A T , BOTMAN C A D , BAKKER N P M
Published: July 2010

Peripheral blood mononuclear cells (PBMC) from Rhesus monkeys previously immunized with bovine type II collagen to induce arthritis were cultured with the same antigen. Because the native protein is poorly soluble in culture medium a healing step is often used. The antigen in this form induced PBMC proliferation, but epitopes for the induction of antibody production and arthritis were lost. To keep the native protein intact it was coated on affigel beads. With the immobilized antigen specific antibody production could be induced.
Predominance of IgGl and IgG4 subclasses of anti-neutrophil cytoplasmic autoantibodies (ANCA) in patients with Wegener's granulomatosis and clinically related disorders
Authors: BROUWER E , TERVAERT J W COHEN , HORST G , HUITEMA M G , VAN DER GIESSEN M , LIMBURG P C , KALLENBERG C G M
Published: July 2010

In view of the supposed hypersensitivity, the elevated levels of IgE, and the occurrence of eosinophilia reported in Wegener's granulomatosis and related conditions, we studied the IgG subclass distribution of ANCA directed against a 29-kD serine protease and myeloperoxidase (MPO) in 41 untreated ANCA-positive patients with several forms of active vasculitis and/or glomerulonephritis. We found that both 29-kD ANCA and MPO ANCA were predominantly of the IgG1 and IgG4 subclass in all groups of patients. The additional presence of IgG3 subclass was associated with renal involvement. We compared the subclass distribution of ANCA with that of total IgG subclass levels, and with the IgG subclass distribution of antibodies to cytomegalovirus (CMV) as a persistent endogenous antigen and antibodies to tetanus toxoid (TT) as an exogenous recall antigen. Total levels of IgG4 were elevated in the majority of the patients together with elevated IgG1 levels. Antibodies to CMV and TT, however, had the same subclass distribution as found in normals and did not show enhanced IgG4 expression. ANCA belong predominantly to the IgG1 and IgG4 subclass, which may suggest that the production of ANCA is related to recurrent exposition to the antigen(s) involved, possibly as part of a hypersensitivity reaction.
Decreased expression of complement receptor type 2 (CR2) on neoplastic B cells of chronic lymphocytic leukaemia
Authors: TOOZE J A , BEVAN D H
Published: July 2010

Neoplastic cells from 49 patients with B cell chronic lymphocytic leukaemia (B-CLL) were studied and compared with normal peripheral and tonsillar B cells using CD2I monoclonal antibodies. Membrane expression of CR2 was quantified by calibrated flow cytometry and by binding analysis with radiolabelled antibody. Both assays indicate that B-CLL cells express only 30% of the CR2 found on normal B cells. These findings are further evidence of the aberrant phenotype of B-CLL cells.
Increased frequency of the null allele at the complement C4b locus in autism
Authors: WARREN R P , SINGH V K , COLE P , ODELL J D , PINGREE C B , WARREN W L , WHITE E
Published: July 2010

Associations between C4 deficiency and autoimmune disorders have been found over the past several years. Since autism has several autoimmune features, the frequencies of null (no protein produced) alleles at the C4A and C4B loci were studied in 19 subjects with autism and their family members. The autistic subjects and their mothers had significantly increased phenotypic frequencies of the C4B null allele (58% in both the autistic subjects and mothers, compared with 27% in control subjects). The siblings of the autistic subjects also had an increased frequency of the C4B null allele, but this increase was not significant. The fathers had normal frequencies of this null allele. All family members had normal frequencies of the C4A null allele, all normal C4A and C4B alleles and all BF and C2 alleles.
Changes in natural killer cell phenotype in patients with post-viral fatigue syndrome
Authors: MORRISON L J A , BEHAN W H M , BEHAN P O
Published: July 2010

We analysed peripheral blood CD56+ natural killer (NK) cell subsets in 23 carefully characterized patients with post-viral fatigue syndrome (PFS), compared with 19 healthy controls, using fluorochrome-conjugated, specific monoclonal antibodies and the FACScan. We found significantly increased percentages of CD56+ and especially CD56bright NK cells in PFS patients. We also found significantly increased percentages of CD56+ high affinity interleukin-2 (IL-2) receptor (CD25)+ and CD56+ transferrin receptor (CD71+) subsets of cells, most of which also stained brightly for CD56. Also, we found an increased percentage of CD56+ CD3+ cells, many of which stained brightly for CD56, although there was no increase in the percentage of CD56- CD3+ T cells in these patients. These observations, in conjunction with very low percentage of CD56- CD25+ cells, suggest that there is a preferential involvement of this minor subset of CD56+ CD3+ T cells in PFS. Finally, a decreased percentage of CD56+ Fc gamma receptor (CD16)+ NK cells was identified, which suggests a reduced capacity for antibody-dependent cellular cytotoxicity in PFS patients. Subsets of CD56+ NK cells co-expressing CD2, CD4 or CD8 did not show any significant difference between PFS patients and healthy controls. These phenotypic changes provide laboratory evidence of immunological abnormalities in this syndrome, and, we suggest, may be consistent with persistent viral infection.
Expression of CD45R0 (UCHL1) by CD4+ and CD8+ T cells as a sign ofin vivoactivation in infectious mononucleosis
Authors: MIYAWAKI T , KASAHARA Y , KANEGANE H , OHTA K , YOKOI T , YACHIE A , TANIGUCHI N
Published: July 2010

CD4SR0 (UCHL1), a member of leucocyte common antigen family, is expressed largely on previously activated or memory T cells. We examined CD45R0 expression of T cell subpopulations in patients with Epstein-Barr virus (EBV) induced infectious mononucleosis (IMN) as a sign of in vivo activation. Consistent with the notion that activated CD8+ T cells expand in acute IMN, the majority of CD8+ T cells in patients with acute IMN expressed CD45R0 to the similar extent to HLA-DR expression. Most CD4+ T cells in these patients also demonstrated marked expression of CD45R0 as well as HLA-DR antigens, compared with age-matched controls. Expression of CD45R0 by CD4+ T cells in patients with acute IMN was more notable than their HLA-DR expression. While predominant CD8+ T cells resulted in decreased percentages of CD4+ T cells, CD4+ T cells expressing CD4SR0 were shown to be significantly elevated in absolute number. The results suggest that both CD4+ and CD8+ T cells may be activated by stimulation with EBV infection. The appearance of two T cell subpopulations expressing CD4SR0 in acute IMN implies their immunoregulatory roles in the control of EBV-infected cells.
Modulation ofMycobacterium aviumgrowthin vivoby cytokines: involvement of tumour necrosis factor in resistance to atypical mycobacteria
Authors: DENIS M
Published: July 2010

The protective mechanisms associated with resistance to atypical mycobacteria infections are not clear. In an effort to broaden our understanding of the mechanisms involved, susceptible mice were infected with a virulent strain of M. avium and various treatments were applied so as to modify the course of the disease. Treatment with an antiserum against tumour necrosis factor-alpha (TNF-α) significantly enhanced the experimental infection, as judged by enumeration of colony-forming units (CPU) in the spleens and livers of infected mice, suggesting a role for TNF-α in resistance to M. avium. In other sets of experiments, recombinant cytokines were directly infused into infected mice. Infusion of recombinant interferon-gamma (IFN-γ) did not modify the experimental infection significantly, and infusion of interleukin-2 was also without effect. Injection of TNF-α enhanced resistance in susceptible animals, as seen by a reduction in the viable bacilli recovered from the spleens and livers. In a final set of experiments, we demonstrate that combinations of cytokines may induce strong resistance against M. avium, namely injection of 1 μg of interleukin-lα and 1 μg of TNF-α at 5-day intervals which was seen to eradicate M. avium in both spleens and livers of susceptible BALB/c mice. Overall, our results suggest that induction of protection against M. avium by treatment with cytokines may be feasible, and that TNF-α may be a pivotal molecule in resistance to M. avium.
Analysis of the human T cell responsiveness to purified antigens ofLeishmania:lipophosphoglycan (LPG) and glycoprotein 63 (gp 63)
Authors: MENDONÇA S C F , RUSSELL D G , COUTINHO S G
Published: July 2010

The responsiveness of peripheral blood lymphocytes from 15 cutaneous leishmaniasis patients and five healthy volunteers to purified surface antigens of Leishmania was studied. The following purified antigens were independently used as stimuli for lymphocyte proliferation in vitro: lipophosphoglycan of L. braziliensis, (Lb-LPG), lipophosphoglycan of L. mexicana (Lm-LPG), glycoprotein 63 of L. braziliensis (Lb-gp 63) and glycoprotein 63 of L. mexicana (Lm-gp 63). Among six patients tested with Lb-gp 63 and Lm-gp 63, two borderline responses and two positive responses were observed, respectively, all the others being negative. Lb-LPG and Lm-LPG induced positive responses in all patients tested (11 with Lb-LPG and five with Lm-LPG). A positive dose-response correlation was observed in the responses of the patients to both Lb-LPG and Lm-LPG. Significant differences (P<0.01) were found between the two groups (patients and healthy subjects) with regard to the responses to Lb-LPG. The proliferating cells after stimulation with Lb-LPG were shown to belong to the CD4+ T cell subset by flow cytometry. However, the following evidence suggests that the T cell responses were not induced by LPG itself but rather by protein contaminants in the LPG preparations: a highly purified Lb-LPG batch that had undergone additional purification induced positive responses in cells from only three out of six patients and a proteinase K-treated Lb-LPG batch did not stimulate any positive response among the same six patients.
Comparison of the T lymphocyte-dependent induction of angiotensin-converting enzyme and leucine aminopeptidase in cultured human monocytes
Authors: ROHRBACH M S , CONRAD A K
Published: July 2010

The T lymphocyte-mediated induction of angiotensin-converting enzyme (ACE) in cultured autologous peripheral blood monocytes has been proposed as a model system for the investigation of the in vivo induction of ACE in the monocyte-derived granuloma epithelioid cells of some granulomatous diseases such as sarcoidosis. The studies described here were designed to evaluate the specificity of the model system by comparing the parameters for induction of ACE with those for the induction of another monocyte metallo-ecto-peptidase, leucine aminopeptidase (LAP). The concentration of LAP in freshly isolated monocytes was 009 mU/106 monocytes (s.e.m. 0.04) and increased to a maximal value of 019 mU/106 monocytes (s.e.m. 0.32) after 3 days when monocytes were cultured alone. ACE was not detectable in freshly isolated monocytes. However, after 6 days of culture, monocytes contained 0.22 m U ACE/106 monocytes (s.e.m. 0.04). Comparison of the levels of ACE and LAP induced during culture of monocytes alone indicated that the induction of these two enzymes was correlated. The induction of both enzymes was further enhanced by the presence of T lymphocytes in a dose-dependent manner. At 4 × 106 T lymphocytes per culture, ACE levels increased to 1.81 mU/106 monocytes (s.e.m. 0.24) and LAP levels to 1.03 mU/106 monocytes (s.e.m. 0.35). The enhancement of ACE activity required autologous lymphocytes, while heterologous T lymphocytes were equally effective in inducing LAP. Comparison of the levels of ACE and LAP induced during co-culture of autologous T lymphocytes and monocytes from 21 independent donors, demonstrated no correlation between the induction of ACE and LAP. These data indicate that, although T lymphocytes also enhance the induction of LAP, the underlying mechanism must differ from that of ACE induction.
A role for prostaglandins in the suppression of cutaneous cellular immunity and tumour development in benzo(a)pyrene- but not dimethylbenz(a)anthracene-treated mice
Authors: ANDREWS F J , HALLIDAY G M , MULLER H K
Published: July 2010

Prostaglandins have been implicated in the immune suppression associated with the development of some tumours. Application of the prostaglandin synthetase inhibitor indomethacin, to murinc skin prior to treatment with the chemical carcinogens benzo(a)pyrene (BP) or7, 12 dimethylbenz(a)anthracene (DMBA), restored contact sensitivity responses to2,4-dinitrofluorobenzene in BP- but not DM BA-treated mice. However, indomethacin failed to restore antibody responses in either group of mice. Prolonged treatment with BP or DMBA led to cutaneous tumour formation. Indomethacin was found to delay the onset and reduce the size of tumours in BP- but not DM BA-treated mice. It is proposed that prostaglandin-induced suppression of cellular cutaneous immunity may play a role in BP- but not DMBA-induced cutaneous carcinogenesis.
Identification of autoantigens in canine autoimmune haemolytic anaemia
Authors: BARKERM R N , GRUFFYDD-JONES T J , STOKEST C R , ELSON C J
Published: July 2010

Autoantigens in canine autoimmune haemolytic anaemia (AIH A) were identified by immunoprecipi-tation using autoantibody eluted from the erythrocytes of affected dogs. At least three patterns of precipitated antigen were identified in six cases of AIH A. The most commonly precipitated antigen pattern was a combination of 42-kD and 29-kD peptides. associated with up to three other membrane components. These autoantigens may be canine glycophorins, which are of similar molecular mass, or may possibly represent an equivalent of the human Rhesus complex. An autoantigen identical in molecular mass to band3, the erythrocyte anion channel protein, was precipitated in one case of AIHA, and unknown peptides of 37 kD and 100 kD were isolated by autoantibody from another dog. In one case, no antigens were precipitated by the eluted antibody, indicating that the autoantibody may have bound a non-protein membrane component such as phospholipid. Overall it is considered that the different patterns observed may reflect differences in the aetiology of the condition. In other studies, sera from dogs with AIHA failed to immunoprecipitate autoantigens, but were shown by immunoblotting to contain autoantibodies to proteins of the erythrocyte cytoskeleton. Such autoantibodies were also demonstrated in normal canine sera and it is suggested that they are unlikely to play a role in the pathogenesis of AIHA, but may be part of a normal clearance mechanism for damaged red blood cells.
Maintained pregnancy levels of oestrogen afford complete protection from post-partum exacerbation of collagen-induced arthritis
Authors: MATTSSON R , MATTSSON A , HOLMDAHL R , WHYTE A , ROOK G A W
Published: July 2010

Pregnancy is known to influence the course of rheumatoid arthritis (RA) in women, as well as type II collagen-induced arthritis (CIA) in DBA/1 mice. A characteristic feature is the remission during gestation and the exacerbation of the diseases during the post-partum period. In the case of CIA in DBA/1 mice, two hormonal changes have been assumed to be critical for the induction of the post-partum flare: (i) the fall in steroid hormone levels from those present during pregnancy; and (ii) surges of prolactin (PRL) release at and after delivery. Our results show that treatment with oestradiol during a short period immediately after parturition protects the mouse from a post-partum flare of the disease, and that treatment with bromocriptine, a drug known to inhibit the endogenous PRL release, has a significant though less marked effect. Studies of lactating (i.e. animals with physiological stimulation of endogenous PRL release) and non-lactating arthritic mice revealed no clear-cut differences, indicating that PRL is of minor importance for the induction of the post-partum flare. Some steroids other than oestradiol, which may be implicated in the exacerbation of arthritis, namely progesterone and hydrocortisone, had no clinical effect, Analyses of agalactosyl IgG levels in mice with CIA, and anti-collagen II antibodies in sera collected at the end of the experiments revealed no significant differences between the oestradiol and the control groups. The successful oestradiol treatment of the mice indicates that the drop in endogenous oestradiol levels prior to delivery ends the oestrogen-mediated protection against arthritis during pregnancy.
Induction of anti-mycobacterial and anti-listerial activity of human monocytes requires different activation signals
Authors: ZERLAUTH G , EIBL M M , MANNHALTER J W
Published: July 2010

The requirements Tor activation of anti-mycobacterial and anti-listerial activity of human monocytes were investigated, Human monocytes could be activated to display enhanced anti-mycobacterial activity by a 24-h treatment with Hpopolysaccharide, The mediator induced by this treatment was identified as being tumour necrosis factor-alpha (TNF-αAQl), Addition of recombinant TNF-α (rTNF-α) to the cultures of human monocytes for 24 h yielded comparable results (minimal dose required for induction of anti-mycobacterial activity, 10 U/ml) Addition of anti-TNF-α antibody completely abrogated the effect, A similar treatment protocol failed to activate enhanced anti-listerial activity, To trigger anti-listerial activity, sequential treatment of human monocytcs with rTNF-α and IL-2 was required, Treatment of monocytes with 10 U/ml rTNF-α for 24 h followed by incubation in the presence of 200 U/ml of IL-2 for an additional 24 h yielded a reduction of listcrial growth which was moderate but statistically significant (P < 0.00l), The activation of monocylcs observed with rTNF-a/IL-2 treatment was (i) dependent on both cytokincs; (ii) sequence dependent (i,e, when IL-2 was added prior to rTNF-α, no effect was observed); and (iii) absent in cells treated with one cytokine only, Enhancement of anti-listerial activity by sequential use of cytokincs was not accompanied by an increase in oxidativc burst, which indicated that oxidative mechanisms were not the reason for the observed Listeria monocytogenes growth restriction, Further support for this hypothesis was obtained after intcrfcron-gamma treatment of human monocytcs which led to an augmented PMA-inducible release of active oxygen radicals, but was not paralleled by growth restriction of L, monocytogenes, Our results indicate that TNI-α plays a crucial role in the activation of monocylcs for growth restriction of intraccllular microbes, Activation of human monocytcs to restrict the growth of the facultative intracellular bacteria Mycohacterium avium intracellulare and L, monocytogenes, however, follows different patterns, the initial trigger in both cases being provided by TNF-α-indueed signals
Immunological tolerance induced by liver grafting in the rat: splenic macrophages and T cells mediate distinct phases of immunosuppressive activity
Authors: YOSHIMURA S , GOTOH S , KAMADA N
Published: July 2010

In the rat combination DA into PVG, liver grafts are not rejected but induce donor-specific transplantation tolerance. We have examined the immunosuppressive properties of spleen cells from PVG recipients of DA liver grafts at various times post-grafting. The results indicate the development of two phases of cell-mediated suppressor activity, which appear to be mediated by separate spleen cell populations. Mitomycin-C-treated spleen cells taken from animals between 5 and 28 days post-grafting were able to suppress rat mixed lymphocyte reactions (MLRs). These ‘early’ suppressor cells were glass adherent and absent from populations purified by passage through nylon wool or G10 Sephadex columns. Suppression of MLR by purified glass adherent cells was not specific for either stimulator or responder haplotypes and was blocked by indomethacin. Nylon wool purified T cells were not suppressive at this time. Spleen cell suppressor activity declined to background levels after 35 days post-grafting. However, spleen cells from long-term surviving liver graft recipients (20 weeks or more) were again able to suppress MLR; the ‘late’ suppressor cells were nylon wool non-adherent and suppression was specific for the donor (DA) MHC type. We conclude that liver grafting in this combination generates early and late phases of suppression among spleen cells, that the early phase is produced by macrophages and mediated by prostaglandins and that the late phase is dependent on allospecific suppressor T cells.
Selective and early increase of IL-1 inhibitors, IL-6 and cortisol after elective surgery
Authors: DI PADOVA F , POZZI C , TONDRE M J , TRITAPEPE R
Published: July 2010

After trauma, inflammatory, immunological and hormonal changes are well documented. Surgical intervention is a form of programmed trauma. Through the study of surgical patients, changes in early endogenous mediators of inflammation, immune response and tissue repair can be investigated. Here we analysed changes in serum levels of IL-1 inhibitors, IL-1β, IL-6, tumour necrosis factor-alpha (TNF-α) and cortisol in patients undergoing elective surgery. C-reactive protein (CRP) was measured as a marker of the acute-phase response. Rises in serum levels of IL-1 inhibitors. IL-6 and cortisol were detected as early as 1 h after the intervention. Peak levels were reached between 2 and 5 h. Serum levels of IL-6 and cortisol remained elevated for several days implying a persistent production. Serum levels of IL-1 and TNF did not change after the intervention. CRP levels peaked on day 2. The communication system sustained by endogenous mediators is activated after surgery as shown by selective changes in IL-1 inhibitors, IL-6 and cortisol. These mediators have different kinetics in serum and IL-6 is not the only early mediator detected. Some IL-1 inhibitors might be involved in the immunological depression observed after major surgery, in the regulation of the inflammatory response or in tissue repair. IL-6 and cortisol seem to act synergistically to activate the acute-phase response. A systemic role for IL-l and TNF is not evident, even if the possibility that these lymphokines may act locally is not ruled out.
Human recombinant erythropoietin directly stimulates B cell immunoglobulin production and proliferation in serum-free medium
Authors: KIMATA H , YOSHIDA A , ISHIOKA C , MASUDA S , SASAKI R , MIKAWA H
Published: July 2010

The effect of human recombinant erythropoietin (Epo) on B cell responses was studied in a serum-free medium. Epo enhanced IgM production and thymidine uptake by a human IgM-producing lymphoblasloid cell line, CBL. This effect was specific to Epo since enhancement was blocked by anti-Epo antibody but not by control antibody. Among the various cytokines, interleukin-4 (IL-4) enhanced IgM production and thymidine uptake while IL-6 enhanced IgM production without affecting thymidine uptake. In contrast, other cytokines including IL-1β, IL-2, IL-5, interferon-alpha (IFN-α), interferon-gamma (IFN-γ), or granulocyte/macrophage colony-stimulating factor (GM-CSF) were without effect. However, the enhancing effect of Epo is different from that of IL-4 or IL-6, since Epo effect was not blocked by anti-IL-4 antibody or anti-IL-6 antibody. Moreover, specific binding of Epo was detected on CBL cells. Epo also enhanced immunoglobulin (IgG, IgM and IgA) production and thymidine uptake by purified tonsil small resting B cells stimulated by Staphylococcus aureus Cowan strain I (SAC) or by large activated B cells. In contrast, Epo had no effect on unstimulated smalt resting B cells. These results indicate that Epo could directly stimulate activated and differentiated B cells and could enhance B cell immunoglobulin production and proliferation.
Regulation of human B cell proliferation and differentiation by seminal plasma
Authors: LEE H-K , LEE H-H , PARK Y-M , LEE J-H , HA T-Y
Published: July 2010

To investigate the role of seminal plasma in human B cell functions, its effect on the proliferation and antibody secretion of tonsillar B cells and an Epstein -Barr virus (EBV) transformed human B cell line, A4, was examined. Seminal plasma inhibited both the proliferation and differentiation of normal B cells only when added to the cultures at the early period of culture. If addition of seminal plasma was delayed beyond 5 to 6 days, it failed to inhibit IgG secretion. Seminal plasma did not show any inhibitory effect on A4 cells, but rather enhanced both the proliferation and IgG secretion of this B cell line. When the low and high mol. wt fractions of seminal plasma were tested for their biological effects on normal and transformed B cells, the low mol. wt fraction (less than 1 kD) was associated with the inhibitory effect of seminal plasma on normal B cells, whereas high mol. wt fractions (both dialysed and 1500-kD fraction) was involved in the enhancing effect on A4 cells. We conclude that (i) seminal plasma inhibits the early proliferation of normal human B cells, but does not inhibit the antibody-secreting capacity of mature B cells; and (ii) different molecules of seminal plasma act on the different stages of B cell maturation.
Defects in proliferative responses of T cells from patients with common variable immunodeficiency on direct activation of protein kinase C
Authors: NORTH M E , WEBSTER A D B , FARRANT J
Published: July 2010

DNA synthesis in response to mitogens has been studied in T cells from nine patients with common variable immunodeficiency (CVI) and seven normal individuals. Five out of the nine patients had cells with subnormal responses to the mitogen phytohaemagglutinin (PHA). As PHA-induced responses are largely mediated through activation of Ca2+-dependent protein kinase C, we studied whether the defective response was still present on direct activation of protein kinase C. This was done using combinations of concentrations of phorbol 12,13,-dibutyrate and the calcium ionophore ionomycin which induced proliferation in normal T cells. We found that in CVI patients with T cells which had normal responses to PHA, responses to phorbol ester and ionomycin were at the same level as in normal T cells. However, with this treatment, in which the linkage between the membrane receptor and protein kinase C is bypassed, the level of DNA synthesis was still depressed in the patient group whose T cells had subnormal responses to PHA. IL-2 failed to restore the DNA synthesis to normal levels when added with the phorbol ester and ionomycin to T cells from one patient in this group. These data suggest that in a group of CVI patients there are defects in T cell activation pathways at or down-stream of protein kinase C.
New Zealand black mice are immunologically resistant to high-dose, but not low-doseLeishmania mexicanainfection
Authors: DOREA R C C , ALEXANDER J , GALLAGHER G
Published: July 2010

The course of infection following s.c. inoculation of a wide dose range of L. mexicana stationary-phase promastigotes (SPP) was examined in sexually mature and immature NZB mice of both sexes. Infection with a high dose (> 107 SPP) was able to induce a protective in vivo response, which could be adoptively transferred with parasite-immune T cells, into naive, syngeneic recipients. In contrast, s.c. infection with a low dose (< 107 SPP) induced non-healing lesions; disease susceptibility could also be transferred into naive animals with T cells from non-immune donors. When the ability to mount a delayed-type hypersensitivity (DTH) reaction was tested in these two groups, the high-parasite dose group gave a significantly higher response. The in vivo protection and high DTH response were reflected in the ability of cells derived from the high-dose resistant (but not low-dose susceptible) mice to mount an antigen-specific T cell response in vitro. The possible immunological effector mechanisms underlying high-dose resistance and low-dose susceptibility are discussed.
Detection of DNA-bound immunoglobulins in patients with lupus nephritis, using monoclonal anti-DNA antibody
Authors: KOYAMA A , KOBAYASHI M , SUZUKI S , SUZUKI M , YAMAGUCHI N , NARITA M
Published: July 2010

In order to investigate the relationship between renal histopathology and the characteristics of circulating immune complexes (CICs) in patients with lupus nephritis (LN), we measured the sizes of CICs, DNA-bound immunoglobulins in patients with systemic lupus erythematosus (SLE) and different histopathological forms of nephritis. Sera were obtained from nine patients: four with diffuse proliferative LN (DPLN), four with membranous LN (MLN), and one with mesangial LN, who fulfilled the criteria of the American Rheumatism Association for SLE. The DNA-bound immunoglobulins were measured by ELISA, in which ELISA plates were coated with mouse monoclonal anti-DNA antibodies. The sizes of CICs were analysed by sucrose density gradient ultracentrifugation. Large (larger than 19S), intermediate (19-7S) and small (nearly 7S) sized DNA-bound immunoglobulins (high peaks of IgG and IgA, but low IgM peaks) were found in the patients with DPLN. By contrast, in patients with MLN, the sizes of ICs; DNA-bound IgG, IgA were in general slightly larger than 7S. In one patient with DPLN, at the onset, various sized DNA-bound IgG, IgA and IgM were found. After the methylprednisolone pulse therapy, CICs became smaller and gradually disappeared. We conclude that the characteristics of DNA-anti-DNA IgG, IgA complexes may determine the localization of ICs in the glomeruli and suggest that CICs play an important role in the pathogenesis of LN.
Serum levels of soluble IL-2 receptor, IL-4 and IgE-binding factors in childhood allergic diseases
Authors: MATSUMOTO T , MIIKE T , YAMAGUCHI K , MURAKAMI M , KAWABE T , YODOI J
Published: July 2010

The serum levels of soluble IL-2 receptor (sIL-2R), IL-4 and IgE-binding factors were examined in children with allergic diseases, and compared with those in non-allergic controls of the same age and sex. The results showed age-related decreases in the serum levels of sIL-2R and IgE-binding factors, but not in that of IL-4 in both allergic and non-allergic individuals. Significant elevation of sIL-2R was observed in sera from children with atopic eczema or history of an anaphylactic reaction to food, as compared with that in non-allergic controls. The serum concentration of IL-4 was elevated in all allergic groups, including cases of atopic eczema, bronchial asthma and anaphylaxis to food, compared with non-allergic controls, and was correlated significantly with the serum level of IgE (r= 0.59). The IgE-binding factor levels in sera from patients aged 6-10 years with bronchial asthma, or patients aged 1-5 years with a history of food anaphylaxis were elevated as compared with those in non-allergic controls of same age. There was no significant correlation between the serum levels of IgE-binding factors and IgE. Since sIL-2R is released by activated T cells, the present study is in favour of T cell activation causing allergic skin disorders. The serum levels of IL-4 as well as IgE did not differ among allergic patients of different clinical categories. The role of IgE in atopic eczema and other allergic diseases is not clearly established; however, it seems likely that IL-4 is deeply involved in the increased production of IgE seen in allergic individuals. The possible involvement of IgE-binding factors in the age-related changes of clinical manifestations in childhood allergic diseases was also discussed.