bluuebunny/crossref_metadata_2025 huggingface dataset:
Humanized monoclonal antibody CAMPATH-1H: myeloma cell expression of genomic constructs, nucleotide sequence of cDNA constructs and comparison of effector mechanisms of myeloma and Chinese hamster ovary cell-derived material
Authors: CROWE J S , HALL V S , SMITH M A , COOPER H J , TITE J P
Published: July 2010

Expression of CAMPATH-1H, a humanized MoAb directed against an abundant surface antigen on human lymphocytes, has been studied using transfected myeloma cells. As the site of chromosome integration of DNA transfected into a cell is random we investigated the feasibility and frequency of hitting a ‘jackpot’ site for expression after co-transfection with CAMPATH-1H heavy and light chain constructs in genomic context. A cell line generating 40 μg/ml of CAMPATH-1H in spent culture supernatant was achieved. The full nucleotide sequence of the CAMPATH-1H heavy and light chain cDNA clones is also shown, in addition a comparison of the effector mechanisms, antibody dependent cellular cytotoxicity and complement dependent cytotoxicity of myeloma cell and Chinese hamster ovary (CHO) cell-derived CAMPATH-1H is reported.
Evidence for prostaglandin-producing suppressor cells in drug-induced liver injury and implications in the diagnosis of drug sensitization
Authors: VICTORINO R M M , MARIA V A J , PINTO L A
Published: July 2010

Lymphocyte reactivity to drugs is present in a majority of cases of drug-induced liver injury when in vitro proliferation assays to the suspected drugs are used. One possible explanation to this could be that adherent suppressor cells mediating their action through the production of prostaglandin E2 would suppress the lymphocyte proliferation to drugs in vitro. We studied 42 patients with a clinical diagnosis of drug-induced liver injury by comparing lymphocyte proliferation observed in cultures with five different concentrations of the suspected drug with the lymphocyte proliferation observed in cultures with drug and a prostaglandin inhibitor (indomethacin). Forty-four healthy subjects and 15 individuals with a recent exposure to the suspected drug without development of adverse drug reactions were also studied as controls. In nine (21%) out of 42 patients with drug-induced liver injury a significant lymphocyte reactivity to drugs was detected. When a prostaglandin inhibitor was added to the cultures, the detection of lymphocyte reactivity increased from 21% lo 57%. No cases of lymphocyte reactivity to drugs or drugs with prostaglandin inhibitor were found in the control groups. The phenomenon of increase of lymphocyte proliferation with the addition of a prostaglandin inhibitor was more frequent in patients whose hepatitis was cured in less than 2 months, was more frequently found in certain pharmacological groups and was significantly associated to a latency period to development of hepatitis of less than 8 days. In conclusion, the in vitro phenomenon described here may be used to improve the ability to demonstrate lymphocyte sensitization in drug-induced liver injury and the clinical correlations found are consistent with the possibility of its relevance in vivo.
In vivoactivation of complement by IgA in a rat model
Authors: STAD R K , BOGERS W M J M , THOOMES-VAN DER SLUYS M E , VAN ES L A , DAHA M R
Published: July 2010

In this study we investigated the capacity of rat IgA to activate complement (C) in vivo rat model. Rat monomeric (m-), dimeric (d-) and polymeric (p-) IgA MoAbs were injected intravenously and assessed for deposition or C3 and C4 on IgA. By ELISA it was shown that both d-and p-IgA bound C3 whereas no binding of C3 by m-IgA was observed. Polymeric IgA was more efficient in binding of C3 as compared with d-IgA. However, in haemolytic assays no consistent decrease of plasma complement levels was observed except for dimeric IgA which induced a marginal consumption of AP50. When rats were pre-treated with cobra venom factor (CVF) to deplete C3, no C3 deposition was found on m-, d- or p-IgA. Neither m- nor d- or p-IgA was able to bind C4 in vivo. In agreement with the results described above, large sized polymeric IgA was shown lobe taken up by Kupffer cells (KC) together with C3. No C3 was detected when rats were depleted of C using CVF. Taken together, the experimental data suggest that d- and p-IgA are able to activate C via the alternative pathway in vivo.
Prevention of spontaneous and cyclophosphamide-induced diabetes in non-obese diabetic (NOD) mice with oral 2-acetyl-4-tetrahydroxybutyIimidazole (THI), a component of caramel colouring III
Authors: MANDEL T E , KOULMANDA M , MACKAY I R
Published: July 2010

The effect of oral administration of THI, a compound present in ammonia caramel food colouring, was studied in spontaneous and induced murine diabetes mellitus. Continuous administration of THI at 400 ppm in drinking water reduced the prevalence of spontaneous diabetes in female NOD/Lt mice from 63% in untreated controls to 8% in treated animals. Since cyclophosphamide (CP) accelerates and intensifies diabetes in NOD mice, we also studied the effect of THI in this model. Diabetes incidence was reduced from 100% in mice given only CP to 13–14% in mice given THI either concurrently or from 14 days previously. Histologically.THI greatly reduced the severity of insulitis. As measured by flow cytometry, all THI-treated mice had a 60–80% reduction in splenic CD4+ and CD8+ T cells. THI-treated mice showed no untoward effects and specifically no weight loss, or pathological changes in their livers, kidneys or lungs. However, there was moderate atrophy of the thymus cortex. THI is a small imidazole-containing compound with structural similarity to histamine and urocanic acid, both known to have immunosuppressive properties. It is a widely used food additive with no known long-term toxic effects al low dosage. Thus, THI could be a useful immunosuppressive agent.
Genital secretory immune response to chronic simian immunodeficiency virus (SIV) infection: a comparison between intravenously and genitally inoculated rhesus macaques
Authors: MILLER C J , KANG D W , MARTHAS M , MOLDOVEANU Z , KIYONO H , MARX P , ELDRIDGE J H , MESTECKY J , MCGHEE J R
Published: July 2010

The humoral and genital secretory immune response to chronic SIV infection was compared between female Rhesus macaques inoculated by i.v. or intravaginal routes. Total IgG levels in serum were 10-fold higher in SIV-infected animals when compared with uninfected controls. Vaginal washes from normal macaques contained predominantly IgA and IgG, while those from SIV-infected animals contained high levels of IgG. The SIV-infected animals had high titres of SIV-specific IgG in serum, with lower but detectable IgA and IgM responses. The genital secretory immune response to SIV was similar in intravenously and intravaginally inoculated animals. The anti-SIV response in the vaginal washes consisted mainly of IgG. Within the lamina propria of the reproductive tract of animals chronically infected with SIV there were essentially no IgA or IgG plasma cells and only a small number of IgM plasma cells, while two normal animals had large numbers of IgA plasma cells. These results suggest that the mucosal immune system of the female reproductive tract is impaired inchronic SIV infection.
Effect of fatty acid anilides on immune responses of Swiss mice
Authors: BLASCO R , RUIZ-GUTIERREZ V
Published: July 2010

The possible relationship between fatty acid anilides and the toxic oil syndrome (TOS) which appeared in Spain in 1981 has been debated during recent years. These anilides have been detected as anomalous compound in toxic oils analysed. After treatment with one daily dose of 50 mg/kg of oleilanilide (88.86% pure) for 5 days, animals showed a tendency towards progressive loss of body weight and a significant increase in serum concentration of immunoglobulins. The percentage of suppressor T cells in spleen diminished significantly compared with the control group. Consequently, an increase in the helper T cells/suppressor T cells was also observed. The production of IgM and IgG in culture was significantly higher than in controls and no differences were seen in IgA synthesis. The functional studies of generation of specific IgM, IgA and IgG suppressor cells at variable doses of concanavalin A (Con A) showed paradoxical behaviour of suppressor T cells generated by low doses of Con A. A similar change occurred at higher doses of Con A. These results suggest that low-dose treatment with oleilanilides induces an alteration in the immune response in Swiss mice.
IL-6 enhances the generation of cytolytic T lymphocytes in the allogeneic mixed leucocyte reaction
Authors: MING J E , STEINMAN R M , GRANELLI-PIPERNO A
Published: July 2010

Cytolytic T lymphocytes (CTL) require soluble proteins termed lymphokines to develop lytic activity. In this report we have studied two of the lymphokines involved in the development of CTL during the allogeneic mixed leucocyte reaction (MLR). High doses of dendritic cells induced lytic activity from purified CD8+ cells in both the murine and human MLR. Under these conditions, IL-2 and IL-6 were endogenously produced and secreted. Antibodies to IL-2 or the IL-2 receptor blocked CTL formation; however, anti-IL-6 receptor antibodies only partially inhibited the response while anti-IL-6 antibodies were largely ineffective. When limiting numbers of antigen-presenting cells were used CTL failed to develop, and neither IL-2 nor IL-6 was secreted into the culture supernatant. Although the addition of IL-6 to such cultures was ineffective in generating CTL, the combination of IL-2 and IL-6 resulted in a 4-5-fold increase in lytic activity over that of IL-2 alone. We conclude that in the allogeneic MLR, IL-2 and IL-6 contribute to the generation of lytically active CD8+ cells, and the effect of IL-6 is evident when the dose of antigen-presenting cell is limited.
Intracellular free Ca2+ fluxes and responses to phorbol ester in T lymphocytes from healthy elderly subjects
Authors: NAYLOR J R , JAMES S R , TREJDOSIEWICZ L K
Published: July 2010

A group of healthy elderly subjects (≥75 years) was selected by the strict criteria of the SENIEUR protocol, and compared with healthy young (≤35 years) volunteers. Mitogenic responses of peripheral blood mononuclear cells to phytohaemagglutinin and anti-CD3 were significantly reduced in the elderly (P <0.0002), thereby confirming that even though in perfect health, elderly individuals show impaired cell-mediated immunity. However, no abnormality of intracellular free Ca2+ fluxes could be detected in purified T cells from the elderly subjects when stimulated with anti-CD3 antibody. Nevertheless, both the proliferative responses of purified T cells to phorbol ester and calcium ionophore (Ionomycin) and the phorbol ester-induced inhibition of the Ca2+ response were defective in the elderly subjects (p <0.003 and p < 0.0002, respectively). These data suggest that signal transduction and the generation of second messengers proceed normally in T cells from the elderly, but downstream events mediated by activation of protein kinase C are dysfunctional.
Antiretroviral treatment reverses HIV-induced reduction in the expression of surface antigens on alveolar macrophages in AIDS patients
Authors: BRAY D H , SQUIRE S B , KAWANA A , JOHNSON M A , POULTER L W
Published: July 2010

MoAbs and immunoperoxidasc methods were used to identify antige-resenting and phagocytic cells and to assess expression of HL-R molecules on cells obtained by bronchoalvcolar lavagc (BAL) from 33 AIDS patients and nine normal volunteers. In 17 patients, not receiving antiretroviral therapy, the expression of HL-R molecules (MoAb RFDR1) as well as the percentages of cells expressing RFD1 marker for antige-resenting cells and RFD7 marker for mature phagocytes were significantly reduced. However, in BAL obtained after commencing treatment with zidovudine (AZT) in 21 patients or with 2′,3′–dideoxyinosine(DD1)in five patients, the expression of the markers studied was found to have returned to levels of expression seen in normal lavages. The changes observed were clearly associated with antiretroviral treatment and did not correlate with applications of other drugs, blood CD4 counts or presence of infectious organisms in BAL fluid. As the alterations in the expression of HLA-DR molecules and RFD1 marker on macrophages have been shown to be associated with functional capacities of these cells, the reversal of impaired expression of phenotypic markers on alveolar macrophages in AIDS patients by AZT and DDI signifies an important ability of these drugs to modify immune reactivity and emphasizes the need to monitor such functions in HIV disease.
Defective expression of CD23 and autocrine growth-stimulation in Epstein-Barr virus (EBV)-transformed B cells from patients with Wiskott-Aldrich syndrome (WAS)
Authors: SIMON H-U , HIGGINS E A , DEMETRIOU M , DATTI A , SIMINOVITCH K A , DENNIS J W
Published: July 2010

WAS is an X-linked, recessive, immune deficiency syndrome, characteristically associated with lymphocyte and platelet dysfunction. Peripheral B lymphocytes from WAS patients are non-responsive to polysaccharidc antigens and show reduced numbers of cells expressing the integral membrane glycoprotein, CD23. The release of CD23 proteolytic fragments, so-called soluble CD23 (sCD23). by B lymphoblasts and EBV-transformed B cell lines has previously been described, and these fragments have been shown to stimulate autocrine growth of these cells. We have found that the surface expression of CD23 is reduced on WAS compared with control EBV-B cells. Surface CD23 levels were reduced two-fold in four WAS cell lines (group I) and nine-fold in four other lines (group II). Group II WAS cell lines also showed reduced growth rates in serum-free medium when compared with group I cell lines and EBV-B cell lines from eight normal subjects. In contrast to the group II WAS lines, group I and EBV-B cells from normal individuals produced an autocrine-growth factor activity which could be absorbed by anti-CD23 antibodies. Immunoprecipitation of sCD23 from culture supernatants confirmed that group I WAS cell lines produced less sCD23, particularly the 37K fragment which was prevalent in control EBV-B cells. Northern analysis showed that CD23 mRNA levels were increased three-fold in group I and unchanged in group II WAS compared with normal EBV-B cell lines, suggesting that decreased surface expression in WAS EBV-B cells reflects post-transcriptional events. Together these results suggest that reduced cell surface expression and aberrant proteolysis of CD23 occurs in WAS patients'B lymphocytes and may contribute to impaired immune function in these patients.
Delineation of pulmonary alveolar macrophage subpopulations by flow cytometry in normal subjects and in patients with lung cancer
Authors: MCDONALD C F , HUTCHINSON P , ATKINS R C
Published: July 2010

We have previously described alterations in pulmonary alveolar macrophage (PAM) function in patients with lung cancer when compared with control subjects. This study examined PAM from five patients with lung cancer, seven normal volunteers and nine control patients, to assess whether any differences in surface phenotypic markers were present in lung cancer versus non-cancer subjects, and what changes might be induced with interferon-gamma (IFN-γ) stimulation. After 3 days'culture with or without IFN-γ no differences were seen in the percentages of cells staining positively in each group for HLA class I. class II and ICAM-1 (CD54) antigens. However, in 13 out of 14 control subjects, and only one out of the five cancer subjects, dual PAM populations were identified. The second PAM population (PAM-2) was larger and demonstrated a higher expression of class I and ICAM-1 antigens. Unlike the unfractionated PAM population, PAM-2 consistently responded to IFN-γ stimulation with an increase in both class I (90 ± 25%) and ICAM-1 (45 ± 10%) antigens, while there was no change in class II antigen expression. In three subjects PAM-2 was found to induce a significantly greater mitogen response than the rest of the PAM population. If confirmed in a larger group of patients, the absence of PAM-2 in the majority of patients with lung cancer may underlie the functional PAM defects observed in these patients.
Nephrotoxic serum nephritis in nude rats: the roles of host immune reactions in the accelerated type
Authors: SATO T , KAWACHI H , MORIOKA T , TAKASHIMA N , SAEKI T , OITE T , SATO N L , TAKEUCHI M , SHIMIZU F
Published: July 2010

By immunization with rabbit immunoglobulins and the injection of a subnephritogenic dose of rabbit nephrotoxic serum (NTS), accelerated-type nephrotoxic serum nephritis (NTN) was induced in heterozygous (rnu/+) rats but not in athymic nude (rnu/rnu) rats. By transferring rat antibody against rabbit immunoglobulins, marked proteinuria was induced also in nude rats (202.0 ± 98.4 mg/ day on day 3) as in rnu/+ rats (122.6 ± 35.3 mg/day on day 3). No marked differences in histological findings could be found between both groups. The most marked increase in the number of intraglomerular infiltrating cells was observed in heterozygous rats indicating that the presence of thymus-derived cells leads to the accumulation of more cells in glomeruli. We conclude that humoral immunity alone is enough to accelerate the pathogenic mechanism which induces glomerular injury with heavy proteinuria in this model.
Interferon-gamma activates the oxidative killing ofCandida albicansby human granulocytes
Authors: STEVENHAGEN A , van FURTH R
Published: July 2010

Although granulocytes are essential for the resistance against infections with Candida albicans, these cells do not kill the ingested yeast optimally. Various cytokines can enhance functional activities of granulocytes. but until now only interferon-gamma (IFN-γ) has been applied more widely, namely in patients with chronic granulomatous disease. Since it is not certain whether IFN-γ is able to enhance the candidacidal activity of granulocytes the present study was undertaken. Human granulocytes incubated with various concentrations of recombinant human IFN-γ (rIFN-γ) were studied for the phagocytosis and intracellular killing of C. albicans and their oxygen metabolism after stimulation with opsonized Candida. Results showed a small increase in the rate of phagocytosis and a dose-dependent increase of the intracellular killing of C. albicans and the production of H2O2. The increased candidacidal activity and H2O2 production by rIFN-γ-stimulated granulocytes were inhibited by diphenylenc iodonium (DPI). From these results it is concluded that the increased candidacidal activity of granulocytes activated by rIFN-γ is caused by the increased production of reactive oxygen radicals.
Association of C4B deficiency (C4B*Q0) with erythema nodosum in leprosy
Authors: de MESSIAS I J T , SANTAMARIA J , BRENDEN M , REIS A , MAUFF G
Published: July 2010

A considerable number of studies have postulated significant associations between susceptibility to the different clinical manifestations of leprosy and the MHC, In this investigation, the association between the MHC class III complement proieins C2, BF, C4A and C4B and leprosy in a patient population of Southern Brazil was studied. A total of 109 non-related leprosy patients was investigated; 73 presented wilh lepromatous leprosy (LL), 46 of Ihem had the immunopathological reaction of erythema nodosum (ENL), the remaining 36 were tuberculoid, borderline and indeterminate leprosy (TIBL) patients. The control group included 172 healthy individuals matched with the patients according lo their ethnic and geographical origin, C2, BF, C4A and C4B allotypes were determined by slandard technologies including Western blots for C2 and C4 variant alleles wilh monoclonal and polyclonal antibodies. Non-expressed (‘silent’) C4 alleles in hemizygously deficient individuals were estimated semiquantitatively on the basis of the C4A and C4B isolype ratio and by the M ASC (‘minimal chi-square’) method. The results showed a significantly elevated presence of the non-expressed C4B allele (C4B*Q0) in the LL and ENL patient groups in comparison with the controls. The most signifieant difference was observed in the ENL group when compared with the controls. In addition, all patients who were homozygously C4B-deficient had ENL, and most of them had the BF*F1 allele. The comparison between LL patients with and without ENL also showed a statistically significant difference in the presence of C4B*Q0, indieating thai C4B deficiency itself is associated with ENL. The relative risk of LL patients with the C4B*Q0 allele suffering from ENL was 53 compared with LL palients without C4B*Q0, Since immune complexes (IC) are considered to be the palhogenic cause of ENL, our findings indicate thai C4B deficieney may play an important role in the abnormal immune response against Myeobaeterium leprae and in the lack of IC clearance, leading to ENL reactions. Individuals wilh this allele seem to be at a higher risk of developing pathologieal immune reactivity in lepromatous leprosy.
Lymphoid cells in afferent and efferent intestinal lymph: lymphocyte subpopulations and cell migration
Authors: ROTHKÖTTER H J , HUBER T , BARMAN N N , PABST R
Published: July 2010

Gut wall emigrating cells have been characterized in the intestinal lymph. The intestinal lymph duct was cannulated in 6-month-old minipigs. Under non-restraining conditions the efferent lymph from the mesenteric lymph nodes was collected in seven normal animals. Lymph coming directly from the guv (afferent lymph) was also collected in 18 pigs after resection of the mesenteric lymph node chains 3 months previously. The intestinal lymph How was similar in both groups (around 18 ml/h). The lymphoid cell yield was 1·2±1·0 ± 106/h in control animals, while in mesenteric lymph node resected pigs it was around 20 times higher (26·2±17·6 × 106/h). In the gut-derived lymph 76·5±8·8% T lymphocytes were observed (CD4+, 48·1±15·5%; CD8+, 53·6±12·7%). The percentage of immunoglobulin-positive cells was lower (IgM+, 10·1±4·5; IgA+, 1·7±1·1). In 14 mesenteric lymph node resected pigs a mean of 5·6±3·1 × 108 lymphocytes from the gut lymph were labelled in vitro with a fluorescent dye and retransfused. The labelling index of fluorescent cells in the intestinal lymph increased rapidly and remained at a high level until 44 h after cell transfusion. A four-to-ten times lower labelling index was found in the spleen, various lymph nodes and Peyer's patches. Most of the recovered lymphocytes were T cells. This model provides access to the cell pool leaving the gut wall, thus allowing an examination of its role in the gastrointestinal tract and other mucosal-lined organs.
Expression of various MHC class II molecules and of intracellular adhesion molecule-1 (ICAM-1) on focal clusters of dendritic cells in iodine deficiency goitres
Authors: DIMAL P , WILDERS-TRUSCHNIG M , MOOIJ P , LEB G , EBER O , LANGSTEGER W , HEBENSTREIT J , BEHAM A , STIEGLER C , DOHR G , DREXHAGE H A
Published: July 2010

Thyroid sections from 18 consecutive euthyroid patients undergoing surgery for iodine deficiency goitre were investigated by means of immunohistochemistry and immunotluorescence. evaluating the expression of MHC class II antigens (HLA-DR, -DP, -DQ, and RFDI) and intercellular adhesion molecule-1 on the formerly described clusters of dendritic cells, as well as on thyrocyles. Eleven of 18 iodine deficiency goitres contained clusters of dendritic cells. These clusters appeared to express only HLA-DR in two cases: in nine of 12 cases they showed a differential expression of class II molecules in the following frequency: HLA-DR > DQ and/or -DP > RFDI. These dendritic cells also were ICAM-1+. In four of 18 iodine deficiency goitres, thyroid epithelial cells showed MHC class II expression in several combinations, but were ICAM-1-. In normal thyroids and in nodular goitres from inhabitants of the endemic area not having an actual iodine deficiency, only sparse clusters of dendritic cells were found; these cells were only HLA-DR+. Follicle lining cells were negative for the MHC class II molecules. In normal thyroids from an area with sufficient iodine supply, no clusters of dendritic cells were seen. The few dendritic cells observed were lying isolated in the interstitium and only positive for H LA-DR and ICAM-1; epithelial cells were negative for the studied markers. These data show clusters of dendritic cells in thyroids of inhabitants of an endemic area. When goitre is accompanied by iodine deficiency at the moment of operation, there appears to be activation of these dendritic cells and of thyroid epithelial cells.
Detection of a circulating form of vascular cell adhesion molecule-1: raised levels in rheumatoid arthritis and systemic lupus erythematosus
Authors: WELLICOME S M , KAPAHI P , MASON J C , LEBRANCHU Y , YARWOOD H , HASKARD D O
Published: July 2010

We have developed a panel of MoAbs against four separate but overlapping epitopes on endothclial cell (EC) vascular cell adhesion molecule-1 (VCAM-1). Two of the MoAbs(1G11 and IE5) inhibited T cell adhesion to tumour necrosis factor (TNF)-activated EC, whilst two MoAbs (1.4C3 and 6D9) did not. Using these MoAbs we have identified a circulating form of VCAM-1 (cVCAM-l) which has identical epitope distribution to the EC form, and which is able to support the adhesion of the human lymphoblastoid cell line Jurkat J6 by a VLA-4- and VCAM-1-dependent mechanism when immobilized from plasma. cVCAM-l isolated by immunoaffinity and size-exclusion chromatographies was shown by SDS-PAGE to have an apparent mol. wt of 85 90 kD. Levels of cVCAM-l were significantly raised (P < 0.001) in rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) compared with normal individuals. It is possible that cVCAM-l may be a useful plasma marker for the diagnosis and management of patients with inflammatory diseases. Furthermore, detection of elevated cVCAM-l levels may act as a guide to the importance of VCAM-1-dependent cell adhesion in different pathological settings.
Divalency of the monoclonal antibody 5-1-6 is required for induction of proteinuria in rats
Authors: NARISAWA M , KAWACHI H , OITE T , SHIMIZU F
Published: July 2010

A single i.v. injection of 3 mg of the F(ab')2 fragment of MoAb 5-1-6 into rats induced immediate proteinuria (128.1 ± 80.7 mg/24 h on day 1) which lasted 1–2 days. In contrast, rats administered 10 mg of the corresponding Fab fragment did not develop abnormal proteinuria even though an equivalent dose of the intact MoAb 5-1-6 far exceeded the nephritogenic dose. The total kidney binding of 125I-Fab fragment was 209.5 ± 34.3 μg/2 kidneys. This exceeded that obtained by injection of 3 mg MoAb 5-1-6 IgG1 (58.9 ± 12.5 μg/2 kidneys at 1 h)and was similar to that obtained following injection of 3 mg F(ab')2 fragment (235.3 ± 16.9 μg/2 kidneys). Immunofluorescence (IF) showed a linear pattern along the glomerular capillary wall at I h after the administration of MoAb 5-1-6 IgG1. F(ab')2 or Fab fragment. On day 5, fine to coarse granules were observed scattered in F(ab')2 injected rat glomeruli. whereas granules were densely localized in Fab-injected rat glomeruli. Complement-depleted rats injected with 3 mg of MoAb 5-1-6 IgG 1 developed proteinuria with the same time course as non-depleted rats. This observation, together with the ability of F(ab')2 to induce proteinuria. indicates that proteinuria induced by MoAb 5-1-6 is complement-independent. This study suggests that MoAb 5-1-6-induced proteinuria is initiated by cross-linking of the epitopes by divalent MoAb 5-1-6 and is independent of complement activity.
The influence of genetic factors on the immune response as judged by pneumococcal vaccination of mono- and dizygotic Caucasian twins
Authors: KONRADSEN H B , HENRICHSEN J , WACHMANN H , HOLM N
Published: July 2010

Eighty-four mono- and dizygotic Caucasian twins randomly chosen, except for age and sex distribution, were vaccinated with a 23-valent pneumococcal vaccine. Blood samples were drawn before and after vaccination and the concentration of IgG. IgG1 and IgG2 pneumococcal antibodies was meaured using an ELISA technique which only detects type-specific capsular antibodies, since C-polysaccharide antibodies in serum were removed. A significantly closer correlation was found regarding mean IgG and IgG2 antibody concentrations after vaccination in mono- compared with dizygotic twins, and this correlation was seemingly type-specific. Since environmental factors in our study population should not contribute more to the immune response in one type of twins compared with the other, we conclude that genetic factors influence the IgG and IgG2 antibody response to pneumococcal vaccination, and that this influence seems to be type-dependent.
The suppressive effect of gelatin-conjugated superoxide dismutase on disease development and severity of collagen-induced arthritis in mice
Authors: KAKIMOTO K , KOJIMA Y , ISHII K , ONOUE K , MAEDA H
Published: July 2010

We studied the effect of superoxide dismutase (SOD) on murine collagen-induced arthritis (CIA), an animal model of human rheumatoid arthritis (RA). Among SOD derivatives studied, only gelatin-SOD conjugate which has prolonged half life in vivo was effective to suppress the development of CIA. while native SOD or gelatin carrier alone was ineffective. Interestingly, pyran polymer-conjugated SOD which also has a long half life showed no suppressive effect on the disease. No significant effect on immune response against type II collagen (CII) was found in any of the experimental groups. In addition, induction of suppressor cells was not detected in spleen or lymph node cells of the gelatin-SOD-treated group. Therefore, these results suggest that oxygen radicals may have an important role in the effector phase of the immune response to manifest this chronic autoimmune polyarthritis. Thus, the use of appropriate antioxidants for the treatment of human RA may be rationalized.
Effect of pregnancy on proteoglycan-induced progressive polyarthritis in BALB/c mice: remission of disease activity
Authors: BUZÁS E I , HOLLÓ K , RUBLICZKY L , GARZÓ M , NYIRKOS P , GLANT T T
Published: July 2010

Proteoglycan-induced arthritis is a murine autoimmune model displaying many similarities to human rheumatoid arthritis and ankylosing spondylitis, as has been documented by clinical, immunological and histopathological studies. Since the onset of arthritis correlates with the serum antibody level to mouse cartilage proteoglycan (PG), it is believed that these autoreactivc antibodies may play crucial roles in the pathological mechanisms of PG-induced arthritis. We have found that fertility in these PG-induced arthritic mice had been reduced but, unlike collagen-induced arthritis, had not been completely lost. Moreover, pregnancy had a beneficial effect upon the clinical symptoms with very little or no influence on scrum antibody levels. Although fertility was retained and arthritic mothers delivered healthy offspring, the birth frequency was significantly less than in non-arthritic age-matched controls. Furthermore, the presence of anti-PG autoantibodies (predominantly IgG1 subclass) transmitted from arthritic mothers to infants transplacentally and by milk during the lactation period did not render these offspring either resistant or more sensitive to subsequent induction of arthritis. Subsequent immunization of infants with ‘arthritogenic’ PG revealed an unaltered susceptibility to arthritis induction.
Complete functional C1q deficiency associated with systemic lupus erythematosus (SLE)
Authors: KIRSCHFINK M , PETRY F , KHIRWADKAR K , WIGAND R , KALTWASSER J P , LOOS M
Published: July 2010

A complete functional deficiency of Clq is described in a patient suffering from SLE. From reduced plasma C1 activity of the parents a hereditary trait was assumed. The defective C1q molecule was haemolytically inactive, did not bind to immune complexes, and was not recognized by the monocyte C1q receptor. C1 activity in the patient's serum could be restored by the addition of purified C1q. Analysis by gelfiltration and ultracentrifugation experiments revealed an immunoreactive molecule of about 150 kD mol. wt, corresponding to one structural subunit of the C1q macromolccule, containing two A chain-B chain dimers and a C-C chain dimer. Applying Southern blot analysis with cDNA clones encoding for the three individual chains of the C1q molecule, no restriction fragment length polymorphism was detected, ruling out possible major alterations of the genetic information.
Factors influencing the occurrence of active cytomegalovirus (CMV) infections after organ transplantation
Authors: BOLAND G J , HENE R J , VERVERS C , De HAAN M A M , De GAST G C
Published: July 2010

In this study, several factors influencing the occurrence of active CMV infection after organ transplantation (Tx) arc analysed. For this purpose, 105 heart, kidney and lung transplant recipients who were CMV-positive or had a CMV-positive donor, were closely monitored for active CMV infection by antigenaemia, cultures. CMV serology and lymphocyte proliferation (LP) to CMV. Univariate and multivariate regression analysis were performed. As pretransplant risk factors the HLA-type and numbers of HLA mismatches between recipients and their donors, and the CMV serology of the recipient and donor were analysed. A new finding was that recipients of donors positive for HLA-B7 were especially at risk for developing active CMV infection (P = 0.03) and CMV disease (P= 0.03). This was not due to increased rejection treatment in these patients. Post-transplant risk factors for development of active CMV infection were absence of detectable cellular immunity to CMV (lymphocyte proliferation) after Tx (P<0.01) and rejection treatment with OKT3 or ATG (P = 0.05). High levels of IgG anti-CMV did not prevent occurrence of active CMV infection or CMV disease in the CMV+ recipients.
Neutrophil activation in ivermectin-treated onchocerciasis patients
Authors: NJOO F L , HACK C E , OOSTING J , STILMA J S , KIJLSTRA A
Published: July 2010

Invermectin is a safe and effective drug for onchocerciasis treatment. In certain individuals, however, therapy is accompanied by adverse reactions. The mechanisms underlying these reactions are not yet known. The aim of the present study was to investigate whether neutrophils are involved in the development of these adverse reactions. Elastase and lactoferrin, two markers for the release of neutrophil azurophilic and specific granule contents respectively, were measured by radioimmunoassays in plasma of onchocerciasis patients with varying degrees of side effects, as well as in control subjects before and 1 and 2 days after ivermectin treatment. A considerable increase of elastase levels after treatment was observed, whereas lactoferrin levels did not change. The percentage of patients with elevated elastase levels was significantly correlated with the degree of side effects. These findings suggest that neutrophil activation may be involved in the development of adverse reactions in these patients.
HLA-B27 subtypes in the spondarthropathies
Authors: MACLEAN I L , IQBALL S , WOO P , KEAT A C S , HUGHES R A , KINGSLEYJ G H , KNIGHT S C
Published: July 2010

The spondartliropathy (Sp)-associated HLA-B27 antigen includes al least seven subtypes. B*2701–07, of which 01, 02, 05 and 07 occur in Caucasians. This study examined the B27 subtype distribution in British patients with Sp. The 133 HLA-B27+ subjecis comprised 94 European Caucasian Sp (58 ankylosing spondylitis (AS), 22 reactive arlhrilis (ReA: 11 sexually acquired (SARA). 11 enteric (EReA)). eight undifferentiated Sp (USp). and six pauciarticular juvenile-onset chronic arthritis (pJCA)) patients, antl 34 healthy Caucasian controls, together with four Asian Indian and one Chinese. 35S-labelled B27 was immunoprecipitated with anti-B27 MoAbs. and subtyped according to isoelectric point (pi) following isoelectric focussing. The use of B27 MoAb permitted subtype assignment without full class 1 HLA lyping. The vast majority (95%) were B*27O5 (Caucasian controls 31/34; AS 55/58; ReA 21/22; USp 8/8. and pJCA 6/6; Indian control 1/1 and AS 2/3: Chinese pJCA I/I), and the remainder B*2702. No B*270l or 07 subjects were identified. AS occurs in both B*2702 and 05 subjects, and we extend this observation to small numbers of ReA and of Indian AS subjects. This implicates molecular features shared between B27 subtypes, rather than subtype-determining regions of the antigen, in Sp palhogenesis.
Augmented lymphocyte binding to cultured endothelium in psoriasis
Authors: LEE M L , TO S S T , COOPER A , JONES M , SCHRIEBER L
Published: July 2010

Lymphocyte binding to cultured human umbilical vein endothelial cells was evaluated using a modified centrifugation binding assay in 15 patients with psoriasis and compared with three patients with atopic dermatitis, 11 patients with rheumatoid arthritis and 28 normal controls. Patients with psoriasis demonstrated 61% augmented lymphocyte binding compared with normal controls (P<0.0001), which was not explained by differences in age and sex or an effect of psoriatic sera. In serial studies of six patients, this difference was found to be reversible with treatment and clinical improvement. Lymphocytes from patients with atopic dermatitis demonstrated decreased binding to endothelium (P<0.005), while those from patients with rheumatoid arthritis were not different from normal controls. This is the first skin disease described in which augmented lymphocyte binding to endothelium occurs, and may represent a mechanism by which lymphocytes are targeted to psoriatic skin.
Early time course of recruitment of immune surveillance in human skin after chemical provocation
Authors: FRIEDMANN P S , STRICKLAND I , MEMON A A , JOHNSON P M
Published: July 2010

As part of the defence function of skin it seems probable that mechanisms exist for the rapid recruitment of immune surveillance to ‘inspect’ any foreign substance that penetrates the skin. In the present study, evidence of such mechanisms was sought by following the time course of early changes in distribution of immune cells, expression of cell adhesion molecules and cytokines after epicutaneous challenge with provoking chemicals to which subjects were known to be either specifically ‘sensitive’ or ‘non-sensitive’; anthralin, an irritant chemical, was used as control. Fiftyseven individuals were studied and there were at least five biopsy samples at each lime point. Regardless of whether individuals were sensitive or not, or of the type of chemical, dermal microvascular endothelial cells showed increased expression of the adhesion molecules ELAM-1 and VCAM-1 within 2 h. and ICAM-I within 8 h. The intensity of immunohistochemical staining increased progressively up to 24 h. More vessels stained for ICAM-1 than for VCAM-1 or ELAM-1. implying that not every vessel expressed all three cell adhesion molecules. Another early change, observed 2 h after irritant challenge, was a significant increase in numbers of CD la+ dendritic cells in the superficial dermis from a median of 3/high power field (hpf) to 9.5/hpf (P<0.03). This was not observed with ‘weak’ provoking substances, such as nickel, but did occur with the potent provoking agent dinitrochlorobenzene (DNCB). Thus, as little as 2 h after contact with provoking chemicals, the skin activates cellular mechanisms to increase T cell infiltration for the presumed purpose of immune surveillance. These mechanisms are not dependent upon specific immune sensitivity and reflect a capacity of skin cells to respond to chemical provocation.
Local immunosuppressive therapy with monoclonal anti-T cell antibody on renal allograft survival in the rat
Authors: LEE C J , YOSHIMURA N , SHIHO O , KITA M , OKA T
Published: July 2010

Considerable interest in the experimental and clinical use of MoAbs as potential therapeutic agents in allograft rejection has been generated by the recent reports of striking prolongation. In this study we investigated the efficacy of the local administration of MoAb OX-19 which is directed to the rat CD5 equivalent, through the renal artery using a rat kidney transplant model, in order to develop a potent method for modifying rejection while minimizing the systemic side effects. Untreated Lewis rats (LEW, RT-11) rejected Brown-Norway rat (BN, RT-1n) kidney at 7·8±0·2 days (n=10). Mean survival time (MST) of recipients treated with OX-19 (75 μg/kg per day) as single bolus injections via the dorsal penile vein for 7 days was 7·0 ± 0·2 days (n= 5, NS). LEW hosts receiving OX-19 (75 μg/kg per day) continuously for 7 days via a femoral vein by using an osmotic minipump (IV-treated group) showed a slight prolongation of graft survival (MST=8·8±0·9 days, n= 5), but this was not statistically significant. On the other hand, local continuous intrarenal arterial infusion of OX-19 (75 μg/kg per day) for 7 days (RA-treated group) significantly prolonged the graft survivals (MST =16·8± 1·3 days, n= 8, P<0·01). Histological examination of MoAb-treated LEW hosts on day 6 post-grafting revealed that kidney grafts from RA-treated hosts showed a slight tubular necrosis, but reduced mononuclear cell infiltration, whereas kidney grafts from IV-treated hosts displayed a severe mononuclear cell infiltration around the artery with interstitial oedema. Moreover, the local intrarenal administration of OX-19, even when the dose is delayed until day 4 after renal grafting, has a therapeutic effect for on-going acute allograft rejection (MST= 11·4 ± 0·8 days, n= 8) compared with administration of OX-19 intravenously from day 4 after grafting (MST= 7·6 ±0·2 days, n= 5,P<0·01) or with no treatment (MST = 7·8±0·2 days, P<0·01). The phenotype of graft infiltrating cells (GIC) was investigated on day 6 post-grafting. There was a significantly lower percentage of cells positive for OX-19, OX-8, OX-26 (transferrin receptor), and OX-39 (1L-2 receptor) in the RA group than in the IV group. These results indicate that local administration of OX-19 via the renal artery is a more efficacious treatment than systemic i.v. injection, and suggests that it may provide a localized immunosuppression of allografts, with reduction of morbidity in clinical transplant recipients.
T lymphocytes in giant cell arteritic lesions are polyclonal cells expressing αβ type antigen receptors and VLA-1 integrin receptors
Authors: SCHAUFELBERGER C , STEMME S , ANDERSSON R , HANSSON G K
Published: July 2010

Giant cell arteritis (GCA) is a common disease in the elderly. It is characterized by focal inflammatory lesions dominated by T lymphocytes and macrophages. The etiology of GCA is, however, still unknown. The aim of the present study was to determine whether lesional T cells represent clonal proliferations, and to characterize adhesion receptors that could be important for recruitment of T cells and antigen receptors involved in their activation. Temporal artery biopsies were obtained from 13 patients presenting with clinical signs of GCA. Immunohistochemistry was used to characterize cell surface receptors on CD3+ T cells in situ in the lesions of eight patients with biopsy-verified GCA. The overwhelming majority of T cells in GCA lesions expressed the TCRγδ receptors. In sections from three of eight patients, a small proportion of cells expressing TCRγδ was also seen. Almost all T cells expressed the integrin receptors, LFA-1 and VLA-1, as determined by double-staining. To characterize the clonal composition of the lesional T cell population, cells were isolated by collagenase digestion of two lesions and T cells cloned by limiting dilution in the presence of mitogenic antibodies, IL-2 and autologous feeder cells. Rearrangements of the T cell receptor (TCR) genes of the clones were analysed by Southern hybridization using probes for TCRγ and β genes. T cell clones established from GCA lesions exhibited heterogeneous rearrangement patterns, indicating a polyclonal origin of the cells. We conclude that GCA lesions contain T lymphocytes that are of polyclonal origin and express integrin-type adhesion receptors. This supports the hypothesis that GCA involves an inflammatory response during which polyclonal T cells adhere to arterial tissue components and accumulate in the developing lesions.
Calreticulin synthetic peptide analogues: anti-peptide antibodies in autoimmune rheumatic diseases
Authors: ROUTSIAS J G , TZIOUFAS A G , SAKARELLOS-DAITSIOTIS M , SAKARELLOS C , MOUTSOPOULOS H M
Published: July 2010

Autoantibodies in sera from patients with systemic lupus erythemalosus (SLE) and onchocerciasis recognize calreticulin (CaR). a calcium-binding protein, as antigen. In this study we present the immunological properties of two synthetic peptides prepared to correspond to the 1–24 and 7–24 amino acid sequence of CaR. In contrast to information previously reported for the recombinant protein, the CaR-peptide analogues appeared immunoreactive to anti-Ro/SSA autoimmune sera. Human sera from patients with SLE, Sjogren's syndrome (SS), rheumatoid arthritis (RA), as well as mixed connective tissue disease (MCTD), demonstrated a positive autoimmune response (binding of antibodies), to the CaR-peptide analogues. These findings suggest that anti-calreticulin autoantibodies are not restricted to any disease specificity.
An improved enrichment method for functionally competent, highly purified peripheral blood dendritic cells and its application to HIV-infected blood samples
Authors: KARHUMAKI E , VILJANEN M E , COTTLER-FOX M , RANKI A , FOX C H , KROHN K J E
Published: July 2010

Dendritic cells (DC) were purified from human peripheral blood using a rapid and simple method based on magnetic depletion of phagocytes with carbonyl iron, followed by centrifugation of non-phagocytic cells on a Percoll density gradient and depletion of lymphocytes and macrophages/ monocytes with a panel of MoAbs and immunomagnetic beads. Enriched DC were obtained with >99% purity as judged by non-specific esterase (NSE) staining. After isolation, these cells, representing 0.4% of the starting mononuclear cell population, still function as potent antigen-presenting cells for purified T lymphocytes. The present results confirm the ability of human peripheral blood DC to present soluble antigens to T cells including microbial antigens and show, further, that DC arc more potent soluble antigen-presenting cells than monocytes. The method was successfully applied to the purification of DC from the blood of HIV-infected individuals. We could not detect decreased numbers of DC in four individuals with early HIV infection and no replicating HIV was detected by in situ hybridization in the DC.
Human recombinant IL-1 receptor antagonist (IL-IRa) inhibits leukotriene B4 generation from human monocyte suspensions stimulated by lipopolysaccharide (LPS)
Authors: CONTI P , PANARA M R , BARBACANE R C , BONGRAZIO M , DEMPSEY R A , REALE M
Published: July 2010

The effect of human recombinant IL-1 receptor antagonist (hrIL-1Ra) on leukotriene B4 (LTB4) release was investigated in activated human monocyte cultures. To stimulate LTB4 generation, LPS was used as an agonist. Detection was performed with the highly sensitive radioimmunoassay method. The cells were treated with scalar concentrations using LPS at 1–1000 ng/ml for different periods of time. The greater LTB4 stimulation was found at LPS 100 ng/ml for 18 h incubation time. Preincubation of monocytes with cytochalasin B (CB) (5 μg/ml) for 15 min augmented the release of LTB4 when LPS was used. A dose-dependent inhibition was found when human monocytes were pretreated for 10 min with hrlL-l Ra at different concentrations (0·25–250 ng/ml) and then treated with LPS 100 ng/ml for 18 h. Maximum inhibition was observed at the highest concentration of hrlL-1 Ra (250 ng/ml). Macrophages treated with a non-selective 5-lipoxygcnasc inhibitor, nordihydro-guaiaretic acid (NDGA), used at 10 μm, added 15 min before LPS 100 ng/ml, produce a dose-dependent inhibition of LTB4. Cells pretreated with arachidonic acid, at various concentrations (10−9− 10−5 m) for 10 min and then treated with LPS 100 ng/ml for 18 h, were also inhibited in a dose-dependent manner by hrIL-1 Ra in their production of LTB4. The inhibition of LTB4 release by hrlL-I Ra, in LPS-stimulated human monocytes, may suggest an important modulatory role for this new cytokine (monokine) in inflammation and immunity and may hold future therapeutic implications for diseases involving LTB4 as a mediator.