bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of ETV6/AML1 abnormalities in acute lymphoblastic leukaemia: incidence, alternative spliced forms and minimal residual disease value
Authors: Codrington Rosalind , O'connor Helen E. , Jalali G. Reza , Carrara Paola , Papaioannou Mary , Hart Stephen M. , Hoffbrand A. Victor , Potter Michael , Prentice H. Grant , Harrison Christine J. , Foroni Letizia
Published: July 2010

The t(12;21)(p13;q22) translocation, resulting in the fusion of the ETV6 and AML1 genes, occurs in 20–25% of paediatric B‐lineage acute lymphoblastic leukaemias (ALL). The identification of the fusion product has important prognostic and therapeutic implications as the translocation has been associated with a favourable clinical outcome. The aim of this study was threefold: (i) to assess the frequency and clinical association of the fusion gene in patients with and without a cytogenetically detectable chromosome 12 and/or 21 abnormality or failed cytogenetic results, (ii) to characterize alternative forms of ETV6/AML1 transcripts, and (iii) to use ETV6/AML1 as a molecular marker for the investigation of minimal residual disease (MRD). ETV6/AML1 fusion was detected in 22 (39%) of 56 cases studied by reverse transcriptase polymerase chain reaction (RT‐PCR). ETV6/AML1 fusion was found in nine out of 16 (56%) cases with a cytogenetically visible chromosome 12 abnormality, but also in nine out of 29 patients (31%) without a chromosome 12 abnormality or patients with failed cytogenetics (four out of 11 patients, 36%), making this the most common cytogenetic abnormality in childhood ALL. Alternatively spliced ETV6/AML1 forms were investigated in 14 of the positive patients. Exon 5 of ETV6 was fused in frame to all AML1 exons, except exon 4. Fusion to exon 6 of AML1 resulted in one amino acid change. The presence of ETV6/AML1 was associated with a lower white blood cell count (Student's t‐test; P = 0·009) and common (c)ALL phenotype (χ2 test; P > 0·001), but no better disease‐free survival. Our study shows that (i) RT‐PCR is the most effective approach for the detection of t(12;21) in childhood ALL, (ii) the association of ETV6/AML1 and chromosome 12 and/or 21, seen in 56% of our cases, further confirms existing data, (iii) overall survival of patients with t(12;21) was not better than other cytogenetics groups, and (d) MRD analysis using ETV6/AML1 fusion is specific, but not sensitive enough to avoid false negative results.
Cytogenetically cryptic AML1–ETO and CBFβ–MYH11 gene rearrangements: incidence in 412 cases of acute myeloid leukaemia
Authors: Rowe D. , Cotterill S. J. , Ross F. M. , Bunyan D. J. , Vickers S. J. , Bryon J. , McMullan D. J. , Griffiths M. J. , Reilly J. T. , Vandenberghe E. A. , Wilson G. , Watmore A. E. , Bown N. P.
Published: July 2010

The rearrangements t(8;21)(q22;22) and inv(16)(p13q22) are two of the most frequently seen in acute myeloid leukaemia (AML), accounting for 8% and 4% of cases respectively. Detection of these abnormalities is important for disease management as both are associated with good responses to conventional chemotherapy and prolonged disease‐free survival. Recent reports using reverse transcriptase polymerase chain reaction (RT‐PCR) suggest that significant proportions of AML cases without a visible t(8;21) or inv(16) show expression of an abnormal fusion gene transcript and, consequently, they could not be detected using conventional cytogenetic analysis alone. We present here a four centre study involving 412 cases of AML screened using both standard cytogenetics and RT‐PCR for AML1–ETO and CBFβ–MYH11. We detected a cytogenetic t(8;21) in 31 out of 412 (7·5%) cases and an inv(16) or t(16;16) variant in 27 out of 412 (6·6%) cases. RT‐PCR detected only two cases (0·5%) of cryptic t(8;21) and no instances of cryptic inv(16). Both cryptic t(8;21) cases had the classic M2 FAB morphology for this type of disease. Our data concur with the established FAB type distribution of the rearrangements and indicate that cryptic t(8;21) and inv(16) may be much less frequent than reported elsewhere.
A comparison of polymorphism in the 3′‐untranslated region of the prothrombin gene between Chinese and Caucasians in Australia
Authors: Chan Daniel K. Y. , Hu Gang , Tao Helen , Owens Daniel , Vun Chee M. , Woo Jean , Chong Beng Hock
Published: July 2010

The 20210G→A mutation in the 3′‐untranslated (UT) region of the prothrombin gene is extremely rare or absent in the Chinese population (0 in 449 subjects, 140 with a history of thromboembolism). This is in contrast to the results from 302 Caucasians from Australia in our study (4·6% in 153 patients with a thromboembolic history and 1·3% in 149 patients with no history). This rarity implies that the variant of the prothrombin gene is probably not the main cause of venous thromboembolism in the Chinese population. Even among Caucasians this mutation accounts for only a minor percentage of all patients with thromboembolism. The relatively low incidence of venous thromboembolism in the Chinese population compared with Caucasians is probably as a result of the low prevalence of factor V Leiden or other environmental or genetic factors.
Natural killer activity and antibody-dependent cellular cytotoxicity in progressive systemic sclerosis
Authors: CIFONE M GRAZIA , GIACOMELLI R , FAMULARO G , PAOLINI R , DANESE C , NAPOLITANO T , PROCOPIO A , PEREGO A M , SANTONI A , TONIETTI G
Published: July 2010

Enhanced natural killer (NK) activity and normal lymphocyte antibody-dependent cellular cytotoxicity (ADCC) were observed in 16 patients with a diagnosis of progressive systemic sclerosis (PSS). Higher NK activity levels were observed against NK-sensitive K562 target cells, while the NK-resistant P815, Daudi and Raji cell lines were not lysed. Cytofluorimetric studies and morphological analysis of peripheral blood lymphocytes (PBL) showed an increased number of CD16 positive cells and large granular lymphocytes (LGL), indicating that the enhancement observed was probably attributable to an increase in the number of circulating NK cells.
Analysis of immunoglobulins secreted by hybridomas derived from rheumatoid synovia
Authors: BROWN C M S , PLATER-ZYBERK C , MAGEED R A , JEFFERIS R , MAINI R N
Published: July 2010

Twenty-six IgG-secreting and eight IgM-secreting hybridomas were derived from the synovia of two patients with rheumatoid arthritis (RA). Hybridomas were obtained by fusing a heteromyeloma cell line, SPAZ-4 with synovial mononuclear cells that were not deliberately stimulated in vitro. Over 96% of the IgG-secreting hybridomas produced antibodies which belonged to the IgGl subclass and showed lambda light chain predominance; the latter was not seen in IgM antibodies, where kappa light chains dominated by 3:1. All IgG antibodies were cationic. Synovial B cells were not exposed to extrinsic stimuli prior to fusion, therefore these results reflect the state of B cell activation and differentiation in vivo. Our results indicate that IgG-secreting B cells in the RA joint are under a selective influence which is, as yet, unidentified. One out of eight IgM-secreting and two out of 26 IgG-secreting hybridomas produced rheumatoid factors (RF). The IgM-RF specificity for IgG heavy chain subclasses was determined and showed that the monoclonal bound to IgG1, IgG2 and IgG4 but not IgG3 with exception of IgG3 Goe of the G3m (st) allotype, a profile typical of specificity for the Ga epitope. This monoclonal also distinguished a determinant in the Fc region of human IgG which was not present in rabbit IgG. The overall frequency of RF-secreting hybridomas we observed indicates that B cells committed to RF production in the synovium of a seropositive and a seronegative RA patient is below 10%.
B cells expressing CD5 are increased in cerebrospinal fluidof patients with multiple sclerosis
Authors: MIX E , OLSSON T , CORREALE J , BAIG S , KOSTULAS V , OLSSON O , LINK H
Published: July 2010

By two-colour flow cytometric analysis, we found increased numbers of B cells co-expressing the pan-T cell marker CD5 and the B cell marker CD19 in cerebrospinal fluid (CSF) of 21 patients withmultiple sclerosis (MS), compared with 17 control subjects with muscular tension headache. Only onepatient with MS, but nine controls lacked CD5+ B cells in CSF. This difference was not observed inperipheral blood. Numbers of CD5+19+ B cells were increased in CSF compared with blood in MS, but not in the controls. In both groups, CD5+19+ B cells were not restricted to small restinglymphocytes, but were also found among larger-sized lymphocytes. The relative density of CD5molecules and of CD19 molecules was lower in CD5+ 19+ thaninCD5- 19+ B cells and CD5+ 19- Tcells. CD5+ B cells are assumed to be responsible for auloanlibody production, and our resultssuggest a palhogenetic role of such cells, predominantly within the central nervous system, in MS.
Cross-reactive rheumatoid factors inrheumatoid arthritis with extra-articular disease
Authors: KINOSHITA M , AOTSUKA S , YOKOHARI R
Published: July 2010

Rheumatoid factors (RF) have been shown to have considerable heterogeneity and bind with IgG aswell as with a variety of substances such as nuclear histone, non-histone nuclear protein, nitrophenylgroups or ssDNA. We describe evidence that polyclonal RF cross-reactive with ssDNA (CRRF) arewidely distributed in a variety of rheumatic diseases, and that their serum level is significantly higherin rheumatoid arthritis (RA) with extra-articular disease. Although the mechanism of the cross-reactivity is not clear, the presence of CRRF could be a serological feature of a clinical subset of RA.The high prevalence rale of CRRF in RA with extra-articular disease also suggests its pathogeneticrole in the extra-articular manifestation of this disease.
Defective interleukin-2 induction of lymphokine-activatedkiller (LAK) activity in peripheral blood T lymphocytesof patients with monoclonal gammopathies
Authors: MASSAIA M , BIANCHI A , DIANZANI U , CAMPONI A , ATTISANO C , BOCCADORO M , PILERI A
Published: July 2010

The recombinant interleukin-2 (rIL-2) generation of lymphokine-activated killer (LAK) cells wasinvestigated in peripheral blood T lymphocytes (PBT) of 16 patients with monoclonal gammopathyof undetermined significance (MGUS) and 32 patients with multiple myeloma (MM). LAK activitywas significantly decreased in MM. but not in MGUS patients, and was partially recovered in MM inthe remission phase. This finding was unexpected, because CD8+ CD11b+ cells, which contain LAKprecursors, are significantly increased in MM. LAK activity was investigated in purifiedCD8+ CD11b+ lymphocytes to discriminate between an intrinsic defect or a defective regulation byother T cell subsets. These cells were intrinsically unable to generate LAK activity fully followingrIL-2 stimulation. MM showed the more pronounced LAK deficiency, while MGUS patients showedintermediate values. Phenotyping revealed significantly increased proportions of Leu7+ and HLA-DR+ cells in MM patients. These data reveal another dysregulation of T cell effector functions inpatients with monoclonal gammopathies and offer further evidence of the impairment of their cell-mediated immunity.
Rapidly proliferative arteriopathy in cyclosporin-induced permanently surviving rat cardiac allografts simulating chronic vascular rejection
Authors: FELLSTRÖM B , DIMÉNY E , LARSSON E , CLAESSON K , TUFVESON G
Published: July 2010

In an attempt to create a model of chronic rejection in rat, cardiac allograft transplantation was performed of hearts from PVG rats to DA rats treated with cyclosporin, 20 mg/kg body weight per day for 14 days and thereafter no immunosuppressive therapy. Experiments were made in 83 rats fed on a normal diet and in 24 rats fed on a diet containing an additional 0·5% cholesterol. Rats on the normal diet showed moderate signs of acute rejection during the first 20–40 days and grafts were lost in acute rejection during this period of time. However, after 2–3 months no signs of acute rejection were present. On the contrary, excessive proliferative changes of the vascular intima and endocardium along with fibrosis and fibrin deposition appeared and was progressive until 6 months post-transplantation. These morphological changes are similar to those found in chronically rejected organs like heart and kidney. In rats fed on a cholesterol diet after cessation of cyclosporin, development of the vascular and endocardial proliferative changes appeared three to four times as fast and were on average fully developed within 4–6 weeks post cessation of cyclosporin treatment. The recipients’ own hearts showed no signs of vascular or endocardial damage. It is thus concluded that two models of vascular and endocardial proliferative changes in cardiac allografts have been developed showing distinct similarities to chronic vascular rejections seen in the clinical transplantation and with apparent similarities to severe arteriosclerosis. The models could be useful in the investigation of pathogenesis and therapeutical means for preventing chronic vascular damage in transplanted organs and arteriosclerosis.
In vitroIgG rheumatoid factor production by CD5-negative murine B cells in response to immune complexes of lipopolysaccharide
Authors: TAGUCHI H , KANOH M , TAKUBO N , KADOTA S , KANAZAWA K , SHIBATA T , UTSUMI S
Published: July 2010

We studied the in vitro production of rheumatoid factor (RF) by spleen cells of normal adult mice. IgG RF cross-reactive with rabbit IgG was produced in response to immune complexes of TNP-lipopolysaccharide (LPS) with murine IgG anti-TNP antibody in an Fc-specific manner, but not to a mixture or IgG and LPS. Antibody-uncomplexed LPS induced little IgG RF production, but suppressed the subsequent IgG RF response to antibody-complexed LPS, whereas IgM RF was induced by either LPS or antibody-complexed LPS. The IgG RF production followed as rapid a time course as IgM RF production; the rate of IgG RF production reached its maximum soon after a lag period of l day and declined after S days. Treatment of splenic B cells from BALBJc mice with anti-Ly-1.2 antibody and rabbit complement resulted in a selective reduction of IgM RF production by 90%, with little effect on IgG RF production. These result suggest that IgG RF is derived primarily from CDS memory B cells which have been developed in normal mice by an unknown mechanism. Unlike the CT)5+ precursor cells for IgM RF, these memory cells arc unresponsive to polyclonal stimulation by LPS but are activated by simultaneous stimulation by aggregated He epilopes and the mitogenic stimulus from LPS.
Cytolytic activity in T cell clones derived from human synovial rheumatoid membrane: inhibition by synovial fluid
Authors: MILTENBURG A M M , VAN LAAR J M , DE KUIPER P , DAHA M R , BREEDVELD F C
Published: July 2010

A panel of T cell clones was derived from the synovial membrane of a patient with rheumatoid arthritis (RA). We investigated whether T cell clones with cytolytic properties were present and whether T cell cytotoxicity was influenced by the presence of synovial fluid. These issues were studied using anti-CD3 and lectin-induced cytotoxicity assays. The majority of the T cell clones derived from the synovial membrane showed cytotoxic properties although non-cytotoxic clones were also found. Three clones (N11, N6 and N15) showed strong cytotoxicity (more than 40% lysis at an effector-to-target cell ratio of 10:1) whereas three clones (N16, N4 and N14) were non-cytotoxic (less than 20% lysis at an effector-to-target cell ratio of 10:1). The induction of cytotoxicity in the anti-CD3-driven system was shown to be dependent on the dose of anti-CD3 present. When synovial fluid was added to these assays a strong inhibition of cytotoxicity was found. This inhibition of cytotoxicity was found with synovial fluid samples of RA patients, as well as with non-RA synovial fluids. Both anti-CD3 and lectin-dependent cytotoxicity assays were strongly inhibited. In conclusion, T cell clones with cytotoxic activity can be isolated from rheumatoid synovial membrane. In the presence of synovial fluid these cytotoxic cells are inhibited to exert their cytotoxic function.
Cytokine regulation of HLA on thyroid epithelial cells
Authors: MIGITA K , EGUCHI K , OTSUBO T , KAWAKAMI A , NAKAO H , UEKI Y , SHIMOMURA C , KURATA A , FUKUDA T , MATSUNAGA M , ISHIKAWA N , ITO K , NAGATAKI S
Published: July 2010

The regulation of class I and class II HLA expression in human thyroid follicular cells was studied in vitro. Tumour necrosis factor-alpha (TNF-α) enhanced the expression of class I antigen on thyrocytes, but these cytokines had little effect on the expression of class II antigen. Interleukin-lβ (IL- lβ) and interleukin-6(IL-6) did not affect class I and class II antigen expression. The combination of interferon-gamma (IFN-γ) with TNF-α or IL-1β enhanced the induction of class I and class II antigens, compared with the effect of IFN-β alone. Neither class I nor class II expression was induced by lL-6 alone or in combination with IFN-γ. These findings suggest that TNF-α and IL-1β may have an important role in inappropriate expression of HLA antigens on thyrocytes in thyroid gland.
Normal and sarcoid alveolar macrophages differ in their ability to present antigen and to cluster with autologous lymphocytes
Authors: GANT V A , SHAKOOR Z , BARBOSA I L , HAMBLIN A S
Published: July 2010

Human bronchoalveolar macrophages from normal individuals function poorly as accessory cells for the presentation of common recall antigens. In sarcoidosis, alveolar macrophages (AM) are reported to be effective accessory cells for the presentation of such antigens. In this study normal and sarcoid AM were compared with blood monocytes for their ability to act as accessory cells in presenting tuberculin purified protein derivative (PPD) and streptokinase-streptodornase (SKSD) to autologous T lymphocytes, or to form spontaneous, antigen- or mitogen-induced clusters with the T cells. When compared to autologous monocytes, normal AM failed to present the two recall antigens effectively. Likewise normal AM formed very few clusters with T lymphocytes when compared to monocytes, even in the presence of antigens or the mitogen phytohaemagglutinin (PHA). In contrast, sarcoid AM presented both antigens as effectively, and were equally effective as monocytes in forming clusters with T lymphocytes, spontaneously and in further response to antigen or mitogen. The results suggest that in sarcoidosis enhanced accessory cell function and enhanced cluster formation may be related features of bronchoalveolar macrophage populations.
The control of class II expression on T cells is independent of the regulation of Tac and the induction of proliferation
Authors: SALVADORI S , PIZZIMENTI A , COHEN S , ZIER K S
Published: July 2010

In order to define the association between class II expression and other markers of T cell activation we tested the ability of various modes of stimulation to induce the expression of class II, Tac, and to stimulate proliferation. Stimulation of T cells with phytohaemagglutinin (PHA) in the presence of accessory cells strongly induced proliferation, Tac and the class II antigen DR. When purified T cells without accessory cells were stimulated with the phorbol ester, PdB, and the calcium ionophore, ionomycin. strong proliferation and Tac expression were induced, but only low levels of surface class II were observed. In contrast, stimulation of the same cells with PHA resulted in weak proliferation, strong Tac, but again low class II levels. The addition of PdB to the PHA increased the proliferative response, but did not affect Tac expression, which remained high, or class II expression, which remained low. Subsequent culture in conditioned medium of purified T cells which had been activated with either PdB and ionomycin or with PHA resulted in increased surface class II levels in both cases. Additional experiments suggested that neither IL-2, IL-4, nor interferon-gamma (IFN-γ) alone was responsible. These results demonstrate that class II expression can be separated from the induction of proliferation and the upregulation of Tac and that the mode of T cell stimulation influences the resulting activation pathway. Furthermore, they suggest that the control of class II expression on T cells is more tightly regulated than it is on other cells.
Hepatitis B surface antigen induces an early-type hypersensitivity
Authors: LEI H Y , WANG J Y , CHANG T T , WANG C C
Published: July 2010

In addition to the delayed-type hypersensitivity (DTH), a unique type of hypersensitivity could be induced at a late stage of the immune responses after hepatitis B surface antigen (HBsAg) immunization. This antigen-specific ear swelling that develops within 1 h after antigen challenge has been referred to as the early-type hypersensitivity (ETH) in contrast to the 24-h DTH. Although expression of ETH was earlier than DTH, the induction of the former needed 3 days longer than that of the latter. In ETH, the plasma protein leaked into the tissue and the vasopermeability increased within 15 min, causing the oedema of ETH. The observation that cyproheptadine, not dexamethasone, inhibited ETH suggests that it is mediated through the release of histamine and/or serotonin. Furthermore, ETH could be transferred by immune sera. Heat treatment (56°C for 4 h) did not destroy the transfer, suggesting that it was not mediated by IgE. The human anti-HBs sera from either hepatitis B virus infection or HBsAg vaccinee also contained the activity to transfer the ETH in mice
Antigen DNA isolated from immune complexes in plasma of patients with systemic lupus erythematosus hybridizes with theEscherichia colilac Z gene
Authors: TERADA K , OKUHARA E , KAWARADA Y
Published: July 2010

Antigen DNA was isolated from immune complexes in plasma of three patients with active systemic lupus erythematosus (SLE) using affinity column. The antigen DNA thus obtained was subjected to hybridization experiments in order to investigate its origin. Unexpectedly, plasmid pUC18s used as a probe was found to hybridize with the antigen DNA. pUC18 was then cleaved into three fragments with the restriction enzyme Haell. A 445-bp fragment containing lac Z DNA hybridized with the antigen DNA. Finally, the lac Z DNA itself was found to hybridize with the antigen DNA. These data strongly suggest that the antigen DNA obtained from three patients is of bacterial origin.
HIV induces modulation of functionally important cellular antigens
Authors: WRIGHTHAM M , SCHIMPF A , PENNINGTON T H , WALKER F , SEWELL H F
Published: July 2010

Infection of T lymphoblastoid CEM cells with the IIIB isolate of HIV-1 results in modulation of the expression of several cellular antigens in addition to the CD4 molecule. The intercellular adhesion receptor LFA-1 (CD 11 a/CD18) and HLA-DR are markedly induced in the cytoplasm and at the cell surface, and the CD7 antigen is down-regulated, being virtually undetectable by sensitive immunocytochemical techniques in the infected cell population. These modulatory effects are to some degree dependent on the virus isolate examined, as the CBL-1 British isolate did not induce comparable phenotypic changes in the CEM cell line. Furthermore, these effects are not reproduced by recombinant gp120 (MB isolate) or p24 added exogenously to uninfected CEM cells. The CD7 molecule appears to play a regulatory role in T cell proliferation, and the LFA-1 integrin molecule is involved in a wide range of immunologically important cell-cell interactions, as well as HIV-induced syncytium formation. The possible contributions of such effects to the pathogenesis of HI V infection are considered.
A 72-kD B cell-associated surface glycoprotein expressed at high levels in hairy cell leukaemia and plasma cell neoplasms
Authors: PULFORD K , MICKLEM K , THOMAS J , JONES M , MASON D Y
Published: July 2010

The present paper describes two new MoAbs, GHI/75 and VMP55, which were raised against a glycoprotein enriched lysate of hairy cell leukaemia. These antibodies recognized a new antigen of 72 kD (unreduced) and 83 kD (reduced) molecular weight. GHI/75 and VMP55 gave very strong staining of plasma cells, moderate labelling of circulating B cells but only weak staining of monocytes, some tissue macrophages and lymphoid cells. Neither antibody reacted with neutrophils or any non-haematopoietic cells. Both antibodies, however, strongly labelled the tumour cells in hairy cell leukaemia, multiple myeloma, plasmacytoma and lymphoplasmacytic lymphomas. No staining was seen of the neoplastic cells in Hodgkin’s disease, myetoid leukaemia or T cell lymphomas. The two antibodies, GHI/75 and VMP55, may be of value in the differential diagnosis of hairy cell leukaemias and plasma cell neoplasms. In addition, the ease with which their antigen can be purified provides the possibility for a detailed study of this molecule.
Pigeon breeders’ lung: pigeon intestinal mucin, an antigen distinct from pigeon IgA
Authors: TODD A , COAN R M , ALLEN A
Published: July 2010

Antigens identified by indirect immunofluorescence staining and specific for sera from patients with pigeon breeders’ lung or healthy pigeon breeders, have been isolated from pigeon intestinal mucus. Two antigenic peaks, one pigeon intestinal mucin and the other IgA, were separated by equilibrium centrifugation of water-soluble mucus in B caesium chloride density gradient. Antigenic positive material was identified by a modified double-sandwich ELISA, by inhibition of haemagglutination of turkey erythrocytes and by gel diffusion. Antigenic-positive material co-fractionated on gel-filtration with purified intact and papain digested pigeon mucin, both free of IgA. These studies demonstrate antibodies to two quite different antigens are associated specifically with sera from pigeon breeders, a novel antigen pigeon intestinal mucin and the previously documented pigeon IgA.
The distribution and functional properties of dendritic cells in patients with seronegative arthritis
Authors: STAGG A J , HARDING B , HUGHES R A , KEAT A , KNIGHT S C
Published: July 2010

Dendritic cells (DC), potent antigen-presenting cells, are known to be increased in numbers in inflammatory lesions in rheumatoid arthritis and juvenile chronic arthritis. In this study, patients with seronegative arthritis were studied: the distribution and functional properties of DC enriched low density cells (LDC) from peripheral blood (PB) and synovial fluid (SF) were compared. The composition of LDC from both sources was similar, comprising approximately 30% DC, 60% monocyles with few T lymphocytes. SF was significantly enriched for LDC compared with paired peripheral blood (P<0.0001) or peripheral blood from healthy controls (P < 0.00l). In contrast, patient PB contained fewer LDC (P < 0.05) overall than healthy controls. LDC from both sources were potent stimulators of allogeneic PB T cells in a mixed leucocyte reaction (MLR), but in four out of 10 patients SF LDC were significantly more stimulatory. In autologous MLRs (AMLRs) SF T cells were not stimulated by cither LDC population. This anergy of T cells was confined to the joint as patient PB T cells showed an AMLR response to PB LDC which was similar to that seen in cells from healthy controls. PB T cells also responded to SF LDC; in a minority of patients SF LDC caused significantly greater stimulation in AMLR than PB LDC and the possibility is discussed that this may represent presentation of antigen acquired in vivo.
Genetic studies on experimental autoimmune gastritis induced by neonatal thymectomy using recombinant inbred strains between a high-incidence strain, BALB/c, and a low-incidence strain, DBA/2
Authors: MORI Y , HOSONO M , MURAKAMI K , KATOH H , YOSHIKAWA Y , KURIBAYASHI K , KANNAGI R , SAKAI M , OKUMA M , MASUDA T
Published: July 2010

Thymectomy on day 3 after birth induced autoimmune gastritis (AIG) at the age of 2 months in 51–73% of BALB/c mice, and in only 3–5% of DBA/2 mice. AIG was detected by histological and serological (immunofluorescence staining for detecting anti-parietal cell autoantibody) examination. However, autoantibody was weakly positive in almost all of these DBA/2 mice when measured by ELISA using extract of murine gastric mucosa as the antigen. To investigate genetically the mechanism controlling the incidence of AIG, 11 recombinant inbred strains established by brother-sister mating of (BALB/c x DBA/2) F2 mice (C x D2 strains) were used. Among 26 markers tested, the Mls-1 locus on BALB/c chromosome 1 and the Hc locus coding a complement component (C5) on BALB/c chromosome 2 were found to be associated with high susceptibility to AIG. However, if one or both of the loci were of DBA/2 origin, mice showed medium or low susceptibility to AIG. For further analysis, F1, F2 and back-cross generations of these two strains were tested, but segregation of a single susceptibility or insusceptibility gene was not obtained. Taken together, it seems probable that two or more genes are involved in the induction mechanism of AIG. We did not detect C5 deposition in AIG lesions, nor complement-dependent cytotoxic antibody to parietal cells in serum from AIG mice. However, injection of irradiated spleen cells of DBA/2 mice into BALB/c mice thymectomized on day 3 augmented the incidence of AIG from 71 to 100%, but not that of oophoritis (33%). A relationship between Mls-1a determinants and the pathogenesis of AIG was further suggested from the fact that Vβ6 TcR-expressing T cells increased in number in AIG-bearing compared with normal BALB/c mice.
Intraperitoneal infection withSalmonella abortusovisis partially controlled by a gene closely linked with theItygene
Authors: OSWALD I P , LANTIER F , MOUTIER R , BERTRAND M-F , SKAMENE E
Published: July 2010

The aim of the present study was to determine whether the Ity gene, which controls the resistance to S. typhimurium infection in mice, also governs the resistance to S. abortusovis, a serotype specific for goat and sheep. During either i.v. or i.p. infection, BALB/c mice (Itys) were not able to control the growth of S. abortusovis and eventually died from infection. In contrast CBA (Ityr) or (C.CB)F1 (Ityr/s) mice were able to control the growth of these bacteria. Using congenic C.D2 Ityr mice, we found that the gene controlling resistance to S. abortusovis was tightly linked to the Ity gene on chromosome 1. Furthermore, in the spleen and the liver of backcross BALB/c × (CBA × BALB/c) mice, the S. abortusovis resistance phenotype cosegregated with the two alleles of the Len-1 gene, a gene tightly linked to the Ity gene. By contrast, in these backcross mice, the level of infection of the peritoneal cavity, the site of inoculation, did not correlate with the Len-1 phenotype of the animal. These results provide evidence that after i.p. inoculation the control of S. abortusovis growth in the spleen and the liver is controlled by the Ity gene, but also suggest that additional gene(s) regulate the number of bacteria at the site of inoculation.
Gut flora induces and maintains resistance against streptococcal cell wall-induced arthritis in F344 rats
Authors: VAN DEN BROEK M F , VAN BRUGGEN M C J , KOOPMAN J P , HAZENBERG M P , VAN DEN BERG W B
Published: July 2010

Streptococcal cell wall (SCW)-induced arthritis is a chronic, erosive polyarthritis that can be induced in susceptible Lewis rats by one i.p. injection of an aqueous, sterile suspension of SCW. F344 rats are resistant to chronic joint inflammation. Our previous studies showed a correlation between susceptibility to SCW-induced arthritis and the ability to mount SCW-specific T cell responses, suggesting tolerance to SCW as a putative mechanism. Here we prevented the induction of tolerance to bacterial epitopes in F344 rats by using them germ-free and analysed susceptibility to arthritis subsequently. In addition, we conventionalized germ-free F344 rats at different times before induction of arthritis. Our results show that germ-free F344 rats are susceptible to SCW-induced arthritis with a similar severity, chronicity, incidence and onset as Lewis rats. Moreover, T cells isolated from germ-free F344 rats were able to respond to SCW. Conventionalization dramatically moderates arthritis and makes T cells unresponsive to SCW again. Thus, in normal rats (F344) a state of tolerance to arthritogenic epitopes is induced (neonatally) and maintained through life by the bacterial flora, resulting in resistance to bacterium-induced artritides. In arthritis-prone (Lewis) rats, this tolerance is deficient and/or easily broken.
IL-2 regulation of soluble IL-2 receptor levels following thermal injury
Authors: TEODORCZYK-INJEYAN J A , SPARKES B G , LALANI S , PETERS W J , MILLS G B
Published: July 2010

In the immunosuppresscd burn patient serum levels of both IL-2 and a soluble form of IL-2 receptor alpha (sIL-2Rα) are significantly elevated. Strikingly, the production of these markers by the in vitro activated patients' cells is decreased. This study examines the role of IL-2 in the decreased production of the sIL-2Rαin vitro in patients with major burns (n = 18, 30 to > 70% total body suface area). Peripheral blood mononuclear cell (PBMC) cultures from patients with highly elevated serum sIL-2Rα. and from healthy controls (n = 12) were activated with concanavalin A (Con A) at initiation. In patients' cultures mitogen-induced increments of sIL-2Rα levels were significantly lower. There was a significant negative correlation (r= −0·64, P<0·001) between a high serum sIL-2Rα level and a decreased Iectin-induced sIL-2Rα release in vitro. Low levels of sIL-2Rα in patients' samples were not normalized by increasing the number of T lymphocytes. Also exogenous rIL-1 was without effect. whereas rIL-3 increased sIL-2Rα release in some cultures. However. sIL-2Rα levels were significantly increased in patients' cultures by (i) addition of exogenous IL-2; (ii) removal of adherent cells; (iii) addition of cyclooxygenase inhibitor, indomethacin: (iv) bypassing cell surface activation by the combination of the calcium ionophore A23187 and the phorbol ester 12-o-tetradreanoyl acetate. The cyclic AMP-elevating drug. forskolin. abrogated the ability of exogenous IL-2 to increase sIL-2Rα production. Thus, in the burn patient, the reduced in vitro sIL-2Rα release appears to relate to abnormalities in IL-2 production and action mediated through its functional surface receptor. Elevated levels of sIL-2Rαin vivo may, therefore, reflect systemic activation of T lymphocytes in response to biologically active IL-2.
Immune defects in families and patients with xeroderma pigmentosum and trichothiodystrophy
Authors: MARIANI E , FACCHINI A , HONORATI M C , LALLI E , BERARDESCA E , GHETTI P , MARINONI S , NUZZO F , RICOTTI G C B ASTALDI , STEFANINI M
Published: July 2010

Xeroderma pigmentosum (XP) is a rare autosomal recessive disease characterized by photosensitivity, a high incidence of cancer in sun-exposed portions of the skin and a reduced capacity to repair the u.v. -induced DNA damage. One of the XP mutations (XP-D) has also been identified in patients affected by trichothiodystrophy (TTD), a rare autosomal recessive disease characterized by brittle hair, mental and physical retardation, peculiar face and ichthyosis. However, in these patients there is no evidence of increased skin tumour incidence. Since an impairment of cell-mediated immunity has been proposed as a co-factor in the cancer proneness of XP patients, we investigated the involvement of immune defect(s) in five XP patients, five TTD patients, their parents, and 24 TTD relatives. We evaluated the phenotype of circulating lymphocytes, natural killer (NK) cell lytic activity, target cell binding of NK cells at single cell level and the effect of interferons (IFN)α and β on NK cell activity. The relative proportion of CD3+ and CD4+ circulating lymphocytes was reduced in XP but not in TTD patients. NK cell lytic activity was decreased in XP patients and their mothers, but their fathers showed normal lytic activity. NK activity varied among TTD families: four out of five patients and their relatives presented low NK cell activity, and one family was normal. In TTD family members, NK activity increased after incubation with IFN-α or IFN-β, but never reached normal values. In contrast, in XP patients and their mothers, the defect was almost completely corrected after in vitro incubation with IFN-α or IFN-β. Our study indicates impaired NK lytic activity in the majority of TTD and XP patients and that this defect is present also in members of their families. In addition, XP patients present a low number of circulating T cells. These multiple abnormalities, together with DNA repair defects, could be related to the increased cancer risk in XP patients.
Reduced PMN β2 integrins after trauma: a possible role for colony-stimulating factors
Authors: WHITE-OWEN C , HARTMANN S , ALEXANDER J W , BABCOCK G F
Published: July 2010

The β2 inlegrins are composed of a common 95-kD β-subunil (CD 18) and one of three possible α-subunils: CDlla, CDllb, or CD1le, These molecules are involved in neulrophil adhesion, diapodesis, chcmotaxis and phagocytosis. In this study, the effects of traumatic injury on neutrophil expression of these α-subunits were investigated. Neutrophils from patients with severe trauma (n = 30) were stained with fluorescent anti-CDlla, -CD1lb, or -CDlle. The percentage of positive neutrophils and the mean channel fluorescence were assayed by flow cytometry. In 10 patients and 10 normals, the effects ol'granulocyte-colony stimulating factor (G-CSF) and granulocyte-maerophage colony-stimulating factor (GM-CSF) on α–subunit expression were evaluated. Ninety-four .2% (s.e.m.) of normal neutrophils were CDlla+, 89.1% were CDllb4 and 89.8% were CDllc+. Only 65 .2% of patient neutrophils were CDlla+, 45.5% wereCDl lb+ and 8.1% were CDllc+ Culture of normal neutrophils without colony-stimulating factors resulted in reduced expression of CDlla and CDllc, but up–regulation of CDllb. Down–regulation of CDlla and CDllc was partially reversed by colony–stimulating factors (30 U/ml). CD1lb receptor density was further up-regulated by GM–CSF and G–CSF. Treatment of patient neutrophils with colony–stimulating factors in culture resulted in up–rcgulation of a–subunils as well. GM–CSF appeared to have the greater effect. These results indicate that colony–stimulating factors may have a clinical role in improving β2 integrin expression, and suggest a use in these infection–prone patients.
Diagnosis of typhoid fever: detection of Salmonella typhi porins-specific antibodies by inhibition ELISA
Authors: NANDAKUMAR K S , PALANIVEL V , MUTHUKKARUPPAN V R
Published: July 2010

Porins are highly immunogenic outer membrane proteins ofSalmonella. Sera from typhoid patients contained a high level of IgG antibodies directed to porins of Salm. typhi. Since porins are highly conserved proteins, anti-porins antibodies both from typhoid patients and healthy normals reacted with porins from several Gram-negative bacteria. Therefore, in order to improve the specificity of detecting Salm. typhi porins-specific antibodies, an inhibition ELISA was developed using enzyme-conjugated MoAbs (MP1 and MPN4) specific to Salm. typhi porins. Sera from typhoid patients with positive haemoculture (16 out of 17) inhibited the binding of MPI to porins, thus showing a positive test for typhoid, whereas sera from patients with other Gram-negative bacterial infections (n= 7) and from healthy volunteers (66 out of 67) were found to be negative. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of this assay were 94.1, 98.7, 97.8, 94.1 and 98.7% respectively. The validity of our inhibition ELISA for typhoid was higher than that of the Widal test. The diagnosis of typhoid fever as early as 3 days after the onset of fever, using a single specimen is possible.
Study of induction of activation of human peripheral blood mononuclear cells with a non-activating form of anti-CD3 MoAb in autoimmune thyroid disease (AITD)
Authors: RESETKOVA E , ARREAZA G , YOSHIKAWA N , MORITA T , KIM H , CARAYON P , VOLPÉ R
Published: July 2010

Anti-CD3 (OKT3) MoAb is a mitogenic agent which activates lymphocytes. We have studied the effects of murine anti-human OKT3 MoAb (IgG1) alone or in combination with IL-2. human thyroglobulin (Tg) and thyroperoxidase (TPO) antigens on the proliferation of whole peripheral blood mononuclear cells (PBMC) (including monocytes) or subtypes (T, CD4+, CD8+, B) as measured by tritiated thymidine (3H-TdR) incorporation. B cell differentiation was studied by measuring numbers of IgG-secreting cells and specific anti-TPO/anti-Tg-secreting cells by SPOT ELISA. PBMC or lymphocyte subtypes, obtained from 45 patients with Hashimoto's thyroiditis (HT). 40 Graves’ disease (GD) and 51 normal controls were cultured in 96 microtitre plates for 6 days in the presence of OKT3 MoAb at final concentrations 25–250 ng/ml, IL-2 15 U/ml. Tg and TPO (I νg/ml). Then cultures were pulsed with 0.2 μCi 3H-TdR/well and incorporation was measured after 18 h. IgG and anti-TPO/Tg-secreting cells were detected at 7 days. Higher proliferative responses from whole PBMC preparations in response to any of the combinations including OKT3 MoAb were observed in the HT preparations, while the basal values were the lowest. IL-2 alone increased these responses markedly, but equally in all groups. IL-2 in combination with OKT3 had an additive effect on proliferation, with higher responses in HT. Tg and TPO antigens did not change these responses. Most HT preparations responded with their maximum proliferation to the lowest concentration of OKT3 MoAb (25 ng/ml), whereas in GD and control preparations of PBMC these responses were shifted to higher concentrations (250 ng/ml); even with those, proliferation was not so enhanced in controls when compared with HT and GD preparations. In contrast, the proliferative responses of T cells alone and subpopulations of CD8+ suppressor/cytotoxic cells were decreased in HT preparations compared with controls. Monocytes were necessary for proliferation. In the subpopulation of B cells (> 95% pure) and CD4+ helper/inducer cells, differences did not reach significance. In spite of the effect on proliferation, OKT3 MoAb only mildly but significantly increased the numbers of IgG-secreting cells in HT and GD preparations and did not stimulate synthesis of specific antibodies. Our data suggest that the increased proliferative responses of whole PBMC to OKT3 MoAb in HT preparations might be due to insufficient activation of T suppressor/cytotoxic cells.
Effect of lipoarabinomannan and mycobacteria on tumour necrosis factor production by different populations of murine macrophages
Authors: BRADBURY M G , MORENO C
Published: July 2010

Tumour necrosis factor (TNF) production is an important pathological mediator in mycobacterial infections, and yet little is known of the factors which influence its production. We have studied the influence of murine macrophage heterogeneity and activation state on TNF production following mycobacterial stimulation in vitro. Lipoarabinomannan (LAM) from strains of Mvcobactcrium tuberculosis and Myco, arium differentially stimulated TNF production in thioglycollate-elicited macrophages in a dose-dependent manner. In comparison, resident peritoneal macrophages produced much less TNF when stimulated with LAM. dead mycobacteria or lipopolysaccharide (LPS). In contrast, zymosan stimulated resident macrophages to a higher degree than thioglycollale-elicited cells. Another comparison between bone marrow and thioglycollalc-elicited macrophages showed that both responded to LPS. but only the latter was stimulated significantly by H37Rv LAM. This may indicate that LAM stimulation of macrophages takes place through a different pathway than both zymosan- and LPS-stimulated TNF production. Also, in vitro activation of peritoneal macrophages with interferon-gamma (IFN-γ), increased TNF response to several stimuli. Our studies indicate that the pathology of mycobacterial infections through TNF production may be influenced by the type and activation state of the macrophage which responds to that infection.
The severe combined immunodeficient (SCID) mouse as a model for the study of autoimmune diseases
Authors: VLADUTIU A O
Published: July 2010

There are no readily available in vivo models to study immune cells from humans with autoimmune diseases. SCID mice, which virtually lack both T and B lymphocytes and accept xenogeneic cells, have been used during the last 5 years to provide a milieu for lymphocytes isolated from individuals with various autoimmune diseases, or for lymphocytes from mice that have a systemic lupus erythematosus-like syndrome. Whilst human autoantibodies to organ antigens have been demonstrated in most SCID mice engrafted with human lymphocytes from the peripheral blood or the target organ, inflammation of the mouse target organ has not generally been observed. This review critically analyses experiments in this area reported so far. Some pitfalls of the SCID mouse model of human autoimmune diseases are mentioned, and future experiments to study mouse and human autoimmunity with this model are proposed.
Phenotypical and functional evaluation of CD8+/S6F1+ T lymphocytes in haemophiliac individuals with HIV-1 infection
Authors: CAVALLIN F , TRALDI A , ZAMBELLO R
Published: July 2010

In this study we investigated the distribution of the S6F1 antigen, an epitope of the lymphocyte function-associated antigen, on CD8+ T lymphocytes in a series of 15 HIV-1+ and 20 HIV-1- haemophiliac patients. MoAbs recognizing the S6K1 antigen have been claimed to distinguish between killer electors (brightly S6F1+ stained) and suppressor cells (dimly S6K1+ stained) within the CD8+ lymphoid population. In addition, we tried to find a correlation between the spontaneous in vitro immunoglobulin synthesis from patients' peripheral blood lymphocytes and the pattern of S6F1 expression. Although the total number of double-positive CD8+/S6F1+ cells was similar in both HIV-1+ and HIV-1- haemophiliac patients, a significant increase in the CD8+/S6F1+ population bright versus dim was documented in HIV-1-infected with respect to HIV-1 haemophiliacs (bright/dim ratio 3·97 ± 0·61 versus 0·75 ± 0·1, respectively, P < 0·005). This finding was correlated to a significant increase in spontaneous in vitro immunoglobulin production in HIV-1+ subjects compared with control haemophiliacs (P < 0·005). Purified CD8+ lymphocytes from HIV-1+ subjects showed a reduced suppressor activity on mitogen-induced immunoglobulin production. Taken together, these data suggest that HIV-1 infection favours the generation of CD8+/S6F1+ bright cells with putative cytotoxic-associated function, leading to a progressive reduction in the number of CD8+/S6F1+ dim suppressor lymphocytes. This phenomenon may contribute to the polyclonal hypergammaglobulinaemia present in HIV-1+ haemophiliac patients.
The induction of cell-associated and secreted IL-1 by iscoms, matrix or micelles in murine splenic cells
Authors: VILLACRES-ERIKSSON M , BERGSTRÖM-MOLLAOGLU M , KÅBERG H , LÖVGREN K , MOREIN B
Published: July 2010

The kinetics of the expression of membrane-associated IL-1 (mIL-1) and soluble IL-1 (sIL-1) was studied in in vitro stimulated spleen cells from non-primed mice or from mice primed with influenza virus antigens incorporated in the immuno-stimulating complexes (iscoms) or as micelles. Matrix, which is the carrier structure for the antigens in the iscom, was used as a non-antigen stimulus. The IL-1 produced was assayed in an IL-1-dependent cell line and the specificity was demonstrated in a blocking experiment with antiserum to IL-1α. Soluble IL-1α was also quantified in ELISA. Iscoms and matrix induced production of mIL-1 and sIL-1 in cultures from non-treated mice as well as from mice primed 4 days before with iscoms or micelles. Micelles were a less strong stimulus and did not induce production of sIL-1. Micelles induced production of mIL-1 in cultures from non-primed mice or from mice which were recently immunized with micelles. No mIL-1 expression was induced by micelles if the spleen cells originated from mice immunized shortly before with iscoms. Depletion experiments demonstrated that sIL-1 was produced by adherent cells upon stimulation with iscoms or matrix. However, factor(s) from the non-adherent cells seem to be necessary for optimal secretion of sIL-1.