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1.
Vascular endothelial growth factor receptor 2 (VEGFR2) has been reported to play an important role in angiogenesis and tumorigenesis. A murine anti-VEGFR2 mAb (A8H1) has been established in a previous study. To reduce the incompatibility of the murine mAb for human use, the chimeric anti-VEGFR2-IgG was developed by genetic recombination of the variable regions of the A8H1 antibody and the constant regions of human IgG, and was expressed in Sp2/0 cells transfected with the two recombinant vectors containing the heavy chain and the light chain regions. After screening, clone 2F12 was selected and was found to stably secrete the murine–human chimeric anti-VEGFR2-IgG (coded 2F12). This chimeric IgG maintained the specificity and the affinity of the parental murine antibody against VEGFR2, and effectively identified VEGFR2 expressed on the surface of HUVECs and BEL-7402 cells. Furthermore, the 2F12 antibody demonstrated inhibition of angiogenesis in vitro, such as proliferation, migration, invasion and tube formation of HUVECs. This murine–human chimeric IgG may be considered for further development as an anti-angiogenesis and anti-tumor agent. 相似文献
2.
Jianfei Huang Yang Tan Qi Tang Xinjian Liu Xiaohong Guan Zhenqing Feng Jin Zhu 《Cytotechnology》2010,62(1):61-71
Vascular endothelial growth factors receptor 2 (VEGFR-2) has been implicated in playing an important role in the formation
of new blood vessels in tumors and other diseases. A high affinity human/mouse cross-reactive anti-VEGFR-2 monoclonal antibody
(mAb) named A8H1 was established by hybridoma technology. Several immunological methods were used to characterize the A8H1,
including ELISA, affinity and kinetics assay, MALDI-TOF MS, WB, IP, IF, FASC and IHC. The results suggested that A8H1 could
bind with linear and conformational epitopes of the VEGFR-2 antigen. The mAb had good specific reactivity with three forms
of VEGFR-2 in HUVEC, and two forms in NIH-3T3 mouse fibroblast cells, which are regarded as non-expressive for VEGFR-2. The
A8H1 mAb associated with intracellular and plasma membranes in HUVEC and with the nuclei in NIH-3T3 cells. This mAb also effectively
identified VEGFR-2 over-expressing cells in a number of archived human cancer tissues. 相似文献
3.
Pharmacokinetics and biodistribution of a light-chain-shuffled CC49 single-chain Fv antibody construct 总被引:5,自引:0,他引:5
Pavlinkova G Colcher D Booth BJ Goel A Batra SK 《Cancer immunology, immunotherapy : CII》2000,49(4-5):267-275
Murine monoclonal antibodies to tumor-associated glycoprotein 72 (anti-TAG-72 mAb B72.3 and CC49) are among the most extensively
studied mAb for immunotherapy of adenocarcinomas. They have been used clinically to localize primary and metastatic tumor
sites; however, murine mAb generally induce potent human anti-(mouse antibody) responses. The immunogenicity of murine mAb
can be minimized by genetic humanization of these antibodies, where non-human regions are replaced by the corresponding human
sequences or complementary determining regions are grafted into the human framework regions. We have developed a humanized
CC49 single-chain antibody construct (hu/muCC49 scFv) by replacing the murine CC49 variable light chain with the human subgroup
IV germline variable light chain (Hum4 VL). The major advantages of scFv molecules are their excellent penetration into the tumor tissue, rapid clearance rate, and
much lower exposure to normal organs, especially bone marrow, than occur with intact antibody. The biochemical properties
of hu/muCC49 scFv were compared to those of the murine CC49 scFv (muCC49 scFv). The association constants (K
a) for hu/muCC49 and muCC49 constructs were 1.1 × 106 M−1 and 1.4 × 106 M−1 respectively. Pharmacokinetic studies in mice showed similar rapid blood and whole-body clearance with a half-life of 6 min
for both scFv. The biodistribution studies demonstrated equivalent tumor targeting to human colon carcinoma xenografts for
muCC49 and hu/muCC49 scFv. These results indicate that the human variable light-chain subgroup IV can be used for the development
of humanized or human immunoglobulin molecules potentially useful in both diagnostic and therapeutic applications with TAG-72-positive
tumors.
Received: 29 December 1999 / Accepted: 4 February 2000 相似文献
4.
Jae‐Hyun Lee Jung‐Ho Sohn Su Yeon Ryu Chein‐Soo Hong Jung‐Won Park 《Journal of cellular and molecular medicine》2013,17(10):1271-1281
Asthma is a chronic inflammatory disease induced by Type 2 helper T cells and eosinophils. Vascular cell adhesion molecule‐1 (VCAM‐1) has been implicated in recruiting eosinophils and lymphocytes to pathological sites in asthma as a regulatory receptor. Accordingly, monoclonal antibody (mAb) against VCAM‐1 may attenuate allergic inflammation and pathophysiological features of asthma. We attempted to evaluate whether a recently developed human anti‐VCAM‐1 mAb can inhibit the pathophysiological features of asthma in a murine asthma model induced by ovalbumin (OVA). Leucocyte adhesion inhibition assay was performed to evaluate the in vitro blocking activity of human anti‐VCAM‐1 mAb. OVA‐sensitized BALB/c mice were treated with human anti‐VCAM‐1 mAb or isotype control Ab before intranasal OVA challenge. We evaluated airway hyperresponsiveness (AHR) and bronchoalveolar lavage fluid analysis, measured inflammatory cytokines and examined histopathological features. The human anti‐VCAM‐1 mAb bound to human and mouse VCAM‐1 molecules and inhibited adhesion of human leucocytes in vitro. AHR and inflammatory cell counts in bronchoalveolar lavage fluid were reduced in mice treated with human anti‐VCAM‐1 mAb as compared with a control Ab. The levels of interleukin (IL)‐5 and IL‐13, as well as transforming growth factor‐β, in lung tissue were decreased in treated mice. Human anti‐VCAM‐1 mAb reduced goblet cell hyperplasia and peribronchial fibrosis. In vivo VCAM‐1 expression decreased in the treated group. In conclusion, human anti‐VCAM‐1 mAb attenuated allergic inflammation and the pathophysiological features of asthma in OVA‐induced murine asthma model. The results suggested that human anti‐VCAM‐1 mAb could potentially be used as an additional anti‐asthma therapeutic medicine. 相似文献
5.
Carla F. M. Molthoff Herbert M. Pinedo Hennie M. M. Schlüper Epie Boven 《Cancer immunology, immunotherapy : CII》1991,34(3):191-197
Summary The new murine anti-episialin monoclonal antibody (mAb) 139H2 has been selected for its strong reactivity with a series of human ovarian cancer xenografts. In the present report we describe the characteristics of mAb 139H2 investigated in vitro as well as in vivo. Scatchard plot analysis using the human ovarian cancer cell line NIH:OVCAR-3 showed an affinity constant of 1 × 108 M–1 and the expression of 7 × 106 antigenic sites/cell. Reactivity with OVCAR-3 xenograft tissue was intense, localized at the cell membrane, heterogeneously distributed, and mainly detectable at the apical site of the cell. Administration of radiolabelled mAb 139H2 to nude mice bearing s.c. OVCAR-3 xenografts showed specific uptake in the tumour up to 9% of the injected dose/g. The maximum uptake in the tumour was retained for 3.5 days and mAb 139H2 cleared from the tumour with a half-life of 5.5 days. The half-life in blood was 50 h and no antibody-antigen complex formation could be detected. Poor uptake and no retention in episialin-negative WiDr colon cancer xenografts demonstrated specificity. Administration of an excess of an unlabelled irrelevant mAb did not influence the uptake in the OVCAR-3 xenografts or in other tissues. In contrast, tumour uptake decreased after addition of 300 µg or more unlabelled mAb 139H2 to a tracer dose of radiolabelled mAb 139H2. The uptake of mAb 139H2 in OVCAR-3 xenografts appeared inversely related to the tumour size.Supported by the Dutch Cancer Society 相似文献
6.
P. K. Wallace Tibor Keler Paul M. Guyre Michael W. Fanger 《Cancer immunology, immunotherapy : CII》1997,45(3-4):137-141
Splenectomy and corticosteroids are the treatment of choice for patients with immune thrombocytopenic purpura (ITP). However,
for the 10%–15% of patients who do not respond to conventional therapy, high-dose i.v. IgG can induce life-saving transient
responses. The benefits of i.v. IgG have been attributed to Fc receptor blockade; however, the involvement of the individual
Fc receptors for IgG (FcγR) in ITP remain to be more completely defined. Recently a mAb, designated mAb H22, which recognizes
an epitope on FcγRI (CD64) outside the ligand-binding domain, was humanized. Because mAb H22 is a human IgG1 and FcγRI has
a high affinity for human IgG1 antibodies, we predicted that mAb H22 would bind to the FcγRI ligand-binding site through its
Fc domain and to its external FcγRI epitope through both Fab domains. These studies demonstrate that mAb H22 blocked FcγRI-mediated
phagocytosis of opsonized red blood cells more effectively than an irrelevant IgG. Moreover, cross-linking FcγRI with mAb
H22 down-modulated FcγRI expression on monocytes, an effect seen within 2 h.
Accepted: 14 October 1997 相似文献
7.
Saha A Baral RN Chatterjee SK Mohanty K Pal S Foon KA Primus FJ Krieg AM Weiner GJ Bhattacharya-Chatterjee M 《Cancer immunology, immunotherapy : CII》2006,55(5):515-527
A murine monoclonal anti-idiotype (Id) antibody, 3H1 has been developed and characterized previously. Anti-Id 3H1 mimics a specific epitope of carcinoembryonic antigen (CEA) and can be used as a surrogate antigen for CEA. 3H1 induced anti-CEA immunity in different species of animals as well as humans and showed promise as a potential vaccine candidate in phase I/II clinical trials for colon cancer patients. One area of interest to us has been the development of new immune adjuvants that may augment the potency of 3H1 as a tumor vaccine. Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are potent immunostimulatory agents capable of enhancing the Ag-specific Th1 response when used as immune adjuvants. In this study, we have evaluated the efficacy of 3H1 as a tumor vaccine when admixed with a select CpG ODN 1826 in transgenic mice that express human CEA. The vaccine potential of 3H1 was also assessed in the presence of another widely used adjuvant, QS-21. 3H1 coupled to keyhole limpet hemocyanin (KLH) and mixed with Freund’s adjuvant (FA) was used as a gold standard in this system. 3H1 vaccination with different adjuvants induced both humoral and cellular anti-3H1, as well as anti-CEA immunity in CEA transgenic mice. The immune sera could lyse CEA-transfected murine colon carcinoma cells, C15 effectively in an antibody-dependent cellular cytotoxicity assay. The anti-CEA antibody responses were somewhat comparable in each adjuvant-treated group of mice, whereas cellular immune responses were significantly greater when CpG was used as an adjuvant. Splenocytes obtained from 3H1–CpG-immunized mice showed an increased proliferative CD4+ Th1-type T-cell response when stimulated in vitro with 3H1 or CEA and secreted elevated levels of Th1 cytokines (IL-2, IFN-γ). This vaccine also induced MHC class I antigen-restricted CD8+ T-cell responses. In a solid tumor model, C15 tumor growth was significantly inhibited by 3H1 vaccinations. In 3H1–CpG-vaccinated mice, the duration of survival was, however, longer compared to the 3H1–QS21-vaccinated mice. These findings suggest that 3H1-CpG vaccinations can break peripheral tolerance to CEA and induce protective antitumor immunity in this murine model transgenic for human CEA. 相似文献
8.
Bufalin, a naturally occurring small-molecule compound from Traditional Chinese Medicine (TCM) Chansu showed inhibitory effects against human prostate, hepatocellular, endometrial and ovarian cancer cells, and leukemia cells.
However, whether or not bufalin has inhibitory activity against the proliferation of human non–small cell lung cancer (NSCLC)
cells is unclear. The aim of this study is to study the effects of bufalin on the proliferation of NSCLC and its molecular
mechanisms of action. The cancer cell proliferation was measured by MTT assay. The apoptosis and cell cycle distribution were
analyzed by flow cytometry. The protein expressions and phosphorylation in the cancer cells were detected by Western blot
analysis. In the present study, we have demonstrated that bufalin suppressed the proliferation of human NSCLC A549 cell line
in time- and dose-dependent manners. Bufalin induced the apoptosis and cell cycle arrest by affecting the protein expressions
of Bcl-2/Bax, cytochrome c, caspase-3, PARP, p53, p21WAF1, cyclinD1, and COX-2 in A549 cells. In addition, bufalin reduced the protein levels of receptor
expressions and/or phosphorylation of VEGFR1, VEGFR2, EGFR and/or c-Met in A549 cells. Furthermore, bufalin inhibited the
protein expressions and phosphorylation of Akt, NF-κB, p44/42 MAPK (ERK1/2) and p38 MAPK in A549 cells. Our results suggest that bufalin inhibits the human lung cancer cell proliferation
via VEGFR1/VEGFR2/EGFR/c-Met–Akt/p44/42/p38-NF-κB signaling pathways; bufalin may have a wide therapeutic and/or adjuvant therapeutic application in the treatment of human
NSCLC. 相似文献
9.
Jie Ma John Samuel Glen S. Kwon Antoine A. Noujaim Ragupathy Madiyalakan 《Cancer immunology, immunotherapy : CII》1998,47(1):13-20
The use of biodegradable poly(dl-lactic-co-glycolic acid) microspheres as a cancer vaccine delivery system for induction of anti-idiotypic responses was investigated
using a murine monoclonal antibody B43.13 that recognizes the human ovarian cancer antigen CA125. Immunization of mice with
mAb B43.13 encapsulated in poly(dl-lactic-co-glycolic acid) microspheres resulted in enhanced humoral and cellular immune responses compared with mAb B43.13 alone or
mAb B43.13 mixed with microspheres. The antibody responses could be further enhanced by the co-encapsulation of mAb B43.13
with monophosphoryl lipid A, a non-toxic adjuvant, in microspheres. Anti-idiotypic humoral responses were shown to result
in Ab2 antibodies mimicking the nominal antigen CA125 and Ab3 antibodies recognizing CA125. Further, microsphere delivery
of mAb B43.13 also resulted in induction of T cell responses involving T2 cells reactive with mAb B43.13 epitopes and T3 cells
recognizing CA125. These results indicate that microsphere delivery of Ab1 can induce both humoral and cellular anti-idiotypic
responses relevant to cancer antigens. This raises the possibility of the use of such formulations for anti-idiotypic induction
immunotherapy for cancer.
Received: 27 August 1997 / Accepted: 24 April 1998 相似文献
10.
Characterization of a human T cell-specific chimeric antibody (CD7) with human constant and mouse variable regions 总被引:2,自引:0,他引:2
G Heinrich H Gram H P Kocher M H Schreier B Ryffel A Akbar P L Amlot G Janossy 《Journal of immunology (Baltimore, Md. : 1950)》1989,143(11):3589-3597
A chimeric human-mouse anti-T lymphocyte mAb (CHT2; SDZ 214-380) has been constructed by cloning the variable region exons of both the L and H chains from the murine hybridoma RFT2 which have CD7 specificity and reactivity with a 40-kDa Ag. The variable L chain exon was joined to the human C kappa, and the variable H chain exon was joined to the human IgG1 region exon encoding the human allotype nGlm(z), nGlm(a). The gene constructs were introduced by electroporation into SP2/0, a non-Ig-producing murine myeloma. The identical tissue reactivity of the newly made CHT2 and the original murine RFT2 mAb (CD7) was confirmed by blocking experiments as well as by immunohistology and flow cytometry. Because this new mAb may have clinical use, the CD7 Ag expression of T lineage cells has also been quantitated in double and triple immunofluorescence assays in combinations with mAb to restricted forms of leukocyte common Ag that designate unprimed (CD45R+) and primed T lymphocyte populations (UCHL1+). CHT2 shows very strong reactivity with large thymic blast cells and cortical thymocytes from which T-ALL originates. Strong staining is seen on CD45R+ unprimed "virgin" T lymphocytes, whereas the expression on UCHL1+ primed "memory" cell types is weaker unless these cells are reactivated by mitogens or Ag. Thus CHT2 may spare a substantial population of resting memory T cells which is relevant to its potential therapeutic use. In addition the chimeric antibody had a greater in vitro antibody dependent cytotoxicity and a prolonged half-life (4.2 to 5.0 days) in Rhesus monkeys. 相似文献
11.
Characterization of a high-affinity monoclonal antibody specific for CD44v6 as candidate for immunotherapy of squamous cell carcinomas 总被引:5,自引:0,他引:5
K.-H. Heider Marlies Sproll Susanne Susani Erik Patzelt Paul Beaumier Elinborg Ostermann Horst Ahorn Günther R. Adolf 《Cancer immunology, immunotherapy : CII》1996,43(4):245-253
Variant isoforms of CD44, a family of cell-surface glycoproteins generated by alternative splicing and post-translational
modifications, are expressed in a variety of human tumors and play important roles in tumor progression and metastasis formation.
The murine monoclonal IgG1 antibody VFF18, specific for an epitope encoded by human CD44 variant exon 6, binds with high affinity
to the recombinant protein (K
d = 1.7×10–10 M) as well as to tumor cell lines in vitro, and is suitable for immunohistochemical analysis of human tumors. Screening of
more than 500 tumor samples of different histogenesis showed that VFF18 most strongly and uniformly reacts with squamous cell
carcinomas (SCC). Detailed analysis of 185 SCC (head and neck, lung, skin) confirmed reactivity of the antibody with 99% of
the samples, with intense and homogeneous staining of the tumor cells in the majority of cases, whereas reactivity of VFF18
with normal tissues is limited to certain epithelia and activated lymphocytes. When radiolabelled VFF18 was administered to
nude mice bearing human epidermoid carcinoma (A-431) xenograft, fast and selective tumor uptake of the radioimmunoconjugate
with a maximum of 18% of the injected dose per gram of tissue was observed. Taken together, these data suggest that mAb VFF18
is a promising targeting vehicle for radioimmunotherapy of squamous cell carcinomas in humans.
Received: 9 September 1996 / Accepted: 25 September 1996 相似文献
12.
A phenyl-beta-galactoside-specific monoclonal antibody reactive with murine and rat NK cells 总被引:1,自引:0,他引:1
V E Miller A E Lagarde B M Longenecker A H Greenberg 《Journal of immunology (Baltimore, Md. : 1950)》1986,136(8):2968-2974
The phenyl-beta-galactoside (phi-beta-gal)-specific monoclonal antibody (mAb) 49H.8 cross-reacts with the terminal disaccharide structure of the asialo GM1 (AGM1) molecule. It was found to react with phi-beta-gal determinants on murine and rat splenic natural killer (NK) cells, as measured by complement depletion studies. Flow cytometric analysis identified the antigen on two IL 2-dependent cloned murine NK cell lines and the rat large granular lymphocyte leukemia RNK. We have compared the 49H.8 reactivity to that of anti-AGM1 antisera (alpha-AGM1) on NK cells and a panel of NK related killer cells, including bone marrow-derived killer cells, lymphokine-activated killer cells (LAK), and anomalous killer cells (AK). We found that the 49H.8 specificity closely paralleled that of alpha-AGM1. When tested against Con A-reactive T cells, the 49H.8 mAb was less reactive than the alpha-AGM1, indicating that it may be a more specific marker for splenic NK populations than the alpha-AGM1. 相似文献
13.
Noboru Oriuchi Naoyuki Watanabe Hidetoshi Kanda Makoto Hashimoto Sumio Sugiyama Seiichi Takenoshita Kyouichi Imai Ryuzou Ueda K. Endo 《Cancer immunology, immunotherapy : CII》1998,46(6):311-317
The study was designed to clarify the difference in pharmacokinetics of monoclonal antibodies (mAb) in animal models and
humans, and to elucidate the applicability of animal models. 99mTc-labeled murine mAb – against carcinoembryonic antigen (designated BW431/26), and neural cell adhesion molecule (NE150)
– and one chimeric mouse/human mAb against nonspecific cross-reacting antigen (chNCA) were administered i.v. to normal mice
and athymic mice (370 kBq, 400 ng) xenografted with human cancer cells expressing antigens, and into patients with tumor (925
MBq, 1 mg). The biodistribution of two of the three mAb (not 99mTc-BW431/26) differed clearly in mice and patients. 99mTc-NE150 showed specific uptake in xenografted tumor and otherwise a normal biodistribution; however, clinical examination
showed increased uptake in the liver with rapid blood clearance (mean α half-life = 31.1 min) compared with 99mTc-BW431/26 (28.4 h). 99mTc-chNCA demonstrated increased blood clearance and renal excretion in both normal and athymic mice, with accumulation in
tumors. Clinical examination showed rapid blood clearance (mean α half-life = 6.4 min) and increased uptake in the liver.
High-performance liquid chromatographic analysis of 99mTc-chNCA revealed the immune complex in blood, suggesting uptake of the complex by the reticuloendothelial cells. The biodistribution
of radiolabeled mAb in animal and human models was variable and specific for each of the three mAb. The results of animal
studies with mAb should be evaluated carefully before being extrapolated to humans, on the basis of the nature of the mAb
and interacting substances.
Received: 9 April 1997 / Accepted 3 March 1998 相似文献
14.
Penichet ML Dela Cruz JS Challita-Eid PM Rosenblatt JD Morrison SL 《Cancer immunology, immunotherapy : CII》2001,49(12):649-662
In the present study we describe a novel murine tumor model in which the highly malignant murine B cell lymphoma 38C13 has
been transduced with the cDNA encoding human tumor-associated antigen HER2/neu. This new cell line (38C13-HER2/neu) showed stable surface expression but not secretion of human HER2/neu. It also maintained expression of the idiotype (Id) of the surface immunoglobulin of 38C13, which serves as another tumor-associated
antigen. Surprisingly, spontaneous tumor regression was observed following s.c. but not i.v. injection of 38C13-HER2/neu cells in immunocompetent syngeneic mice. Regression was more frequently observed with larger tumor cell challenges and was
mediated through immunological mechanisms because it was not observed in syngeneic immunodeficient mice. Mice that showed
complete tumor regression were immune to challenge with the parental cell line 38C13 and V1, a variant of 38C13 that does
not express the Id. Immunity could be transferred with sera, suggesting that an antibody response mediated rejection and immunity.
Continuously growing s.c. tumors as well as metastatic tumors obtained after the i.v. injection of 38C13-HER2/neu maintained expression of human HER2/neu, which can serve as a target for active immunotherapy. As spontaneous tumor regression has not been observed in other human
murine models expressing human HER2/neu, our results illustrate the enormous differences that can exist among different murine tumors expressing the same antigen.
The present model provides a useful tool for the study of the mechanisms of protective immunity to B cell lymphoma and for
the evaluation of different therapeutic approaches based on the stimulation or suppression of the immune response.
Received: 2 August 2000 / Accepted: 20 September 2000 相似文献
15.
K. Kissel S. Hamm Martina Schulz Annunciata Vecchi Cecilia Garlanda B. Engelhardt 《Histochemistry and cell biology》1998,110(1):63-72
A novel monoclonal antibody (mAb), 8D3 (IgG2a), that specifically recognizes the murine transferrin receptor (TfR) was produced
by immunizing a Lewis rat with a polyoma middle T oncogene-transformed endothelioma cell line. The 8D3 mAb was obtained by
immunohistochemical screening for exclusive staining of vessels forming a blood–brain barrier (BBB), but not of other vessels.
The anti-TfR mAb 8D3 recognizes the TfR also in FACS analysis and in western blots and should prove to be useful for affinity
purification of the TfR. Whereas 8D3 brightly stains BBB-forming vessels in the central nervous system of mice, it does not
stain the fenestrated capillaries within the choroid plexus and the circumventricular organs. In testis, where the blood–tissue
barrier is located at the level of the Sertoli cells, the 8D3 mAb specifically stains Sertoli cells but not endothelial cells.
Finally, in vitro, 8D3 does not interfere with iron uptake of lymphocytes as it does not influence their proliferation. Taken
together, 8D3 represents a versatile new tool to study the tissue distribution of the murine TfR and TfR-mediated transcytosis
across tissue barriers in the mouse.
Accepted: 7 January 1998 相似文献
16.
Changes in the levels of antiapoptotic protein B-cell lymphoma 2 (Bcl-2) protein has been reported in murine and human tuberculosis.
We investigated the role of mitogen-activated protein kinase pathways in the production of Bcl-2 protein in THP-1 human monocytes
infected with Mycobacterium tuberculosis H37Rv and H37Ra. Analysis of phosphorylation profiles of mitogen-activated protein kinase kinase-1, extracellular-signal
regulated kinase 1/2, mitogen-activated protein kinase kinase 3/6, and p38 mitogen-activated protein kinase; B-cell lymphoma
2 kinetics; and tumor necrosis factor-α (TNF-α) secretion levels showed variation between the two strains. Mycobacterium tuberculosis H37Rv induced higher Bcl-2 and lower TNF-α levels, whereas H37Ra the reverse. The strains also differed in their usage of
CD14 and human leukocyte antigen-DR receptors in mediating extracellular-signal regulated kinase 1/2 and p38 mitogen-activated
protein kinase activation. Mycobacterium tuberculosis H37Rv- and H37Ra-induced Bcl-2 production was reduced by specific inhibitors of mitogen-activated protein kinase kinase-1
(PD98059) and p38 (SB203580), but increased by nuclear factor κB (NF-κB) inhibitor (BAY 11-7082). TNF-α production by both
strains was reduced in the presence of specific inhibitors of mitogen-activated protein kinase kinase-1 (PD98059), p38 (SB203580),
and NF-κB (BAY 11-7082). Furthermore, inhibition of NF-κB was accompanied by an increase in strain-induced extracellular-signal
regulated kinase 1/2 phosphorylation. Collectively, these results indicate for the first time that the production of Bcl-2
and TNF-α by M. tuberculosis H37Rv/H37Ra-infected THP-1 human monocytes is mediated through mitogen-activated protein kinases and NF-κB. 相似文献
17.
A novel monoclonal antibody against murine IL-2 receptor beta-chain. Characterization of receptor expression in normal lymphoid cells and EL-4 cells 总被引:17,自引:0,他引:17
T Tanaka M Tsudo H Karasuyama F Kitamura T Kono M Hatakeyama T Taniguchi M Miyasaka 《Journal of immunology (Baltimore, Md. : 1950)》1991,147(7):2222-2228
A mAb specific for the murine IL-2R beta-chain (IL-2R beta) was produced by immunizing a rat with a rat transfectant cell line expressing a large number of cDNA-encoded murine IL-2R beta. The mAb, designated TM-beta 1, is specifically reactive with the murine IL-2R beta cDNA-transfectant but not with the recipient cell, and immunoprecipitates murine IL-2R beta of Mr 75 to 85 kDa. TM-beta 1 mAb completely abolished the high affinity IL-2 binding by inhibiting the ligand binding to IL-2R beta. Murine IL-2R beta was found to be constitutively expressed on a subpopulation of CD8+ T cells and almost all NK1.1+ NK cells in the spleen, whereas TM-beta 1 mAb inhibited the proliferation of spleen cells induced by 1 nM of IL-2. Interestingly, EL-4 cells that express murine IL-2R beta as detected by TM-beta 1 mAb can bind neither human nor murine IL-2 under the intermediate affinity conditions, although cDNA-directed human IL-2R beta expressed in the same EL-4 cells has been previously shown to manifest the intermediate affinity IL-2 binding. These results may imply that functional expression of IL-2R beta is differentially regulated between humans and mice. Finally, our neutralizing anti-IL-2R beta mAb TM-beta 1 will be useful not only for various in vitro studies but also for in vivo studies to directly investigate the possible involvement of the IL-2/IL-2R pathway in the generation and differentiation of T lymphocytes and NK cells. 相似文献
18.
Flieger D Kufer P Beier I Sauerbruch T Schmidt-Wolf IG 《Cancer immunology, immunotherapy : CII》2000,49(8):441-448
Cytokine-induced killer cells (CIK), generated in vitro from peripheral blood mononuclear cells (PBMC) by addition of interferon
γ (IFNγ), interleukin-2 (IL-2), IL-1 and a monoclonal antibody (mAb) against CD3, are highly efficient cytotoxic effector
cells with the CD3+CD56+ phenotype. In this study, we evaluated whether the cytotoxicity of these natural-killer-like T lymphocytes against the colorectal
tumor cell line HT29 can be enhanced by the addition of a bispecific single-chain antibody (bsAb) directed against EpCAM/CD3.
For determination of bsAb-redirected cellular cytotoxicity we used a new flow-cytometric assay, which directly counts viable
tumor cells and can assess long-term cytotoxicity. We found that this bsAb induced distinct cytotoxicity at a concentration
above 100 ng/ml with both PBMC and CIK at an effector-to-target cell ratio as low as 1:1. CIK cells revealed higher bsAb-redirected
cytotoxicity than PBMC. Cellular cytotoxicity appeared after 24 h whereas PBMC showed the highest bsAb-redirected cytotoxicity
after 72 h. The addition of the cytokines IL-2 and IFNα but not granulocyte/macrophage-colony-stimulating factor enhanced
bsAb-redirected cytotoxicity of both PBMC and CIK. When the bsAb was combined with the murine mAb BR55-2, which recognizes
the Lewisy antigen, bsAb-redirected cytotoxicity was partly augmented, whereas murine mAb 17-1A, which binds to EpCAM as well, slightly
suppressed bsAb-redirected cytotoxicity induced by the bsAb. We conclude that CIK generated in vitro or in vivo combined with
this new EpCAM/CD3 bsAb and the cytokine IL-2 should be evaluated for the treatment of EpCAM-expressing tumors.
Received: 9 December 1999 / Accepted: 18 May 2000 相似文献
19.
H Crespeau C Alliot J M Fine D Rochu 《Comptes rendus de l'Académie des sciences. Série III, Sciences de la vie》1991,313(7):287-292
The diversity of a polyclonal anti-idiotypic response (Ab2) to a murine monoclonal anti-A (Ab1) was investigated after purification of two Ab2 populations. One was eluted from human polyclonal anti-A column and the other from Ab1. Analysis of the Ab specificity, as well as screening of the clonotypic distribution, were achieved after splitting Ab by IEF; this was followed by immunoblotting and probing with various anti-ABH mAb. The first population reacted with almost all the murine anti-ABH mAb, as well as with four human anti-A mAb, and consequently consisted of Ab2 beta. The second was composed of "true" Ab2 directed against Ab1. In the first population internal images mimicked either A Ag, or H Ag, or some epitopes common to both. This study demonstrates the plurality of internal images-bearing Ig molecules, some mimicking completely, and some only partially or even unfaithfully the nominal A determinant. The analysis of this idiotypic cascade proves the existence of a degeneracy of the initial restricted antigenic specificity. The consequences of such a process are discussed. 相似文献
20.
Xiaoen Wang Andrea J. Bullock Liang Zhang Lin Wei Dongyin Yu Kedar Mahagaokar David C. Alsop James W. Mier Michael B. Atkins Angela Coxon Jon Oliner Rupal S. Bhatt 《Translational oncology》2014,7(2):188-195
Angiopoietin 2 (Ang2) is a secreted glycoprotein upregulated at sites of angiogenesis and has been implicated in cancer neovascularization. Recent studies have suggested efficacy of combined Ang and vascular endothelial growth factor receptor (VEGFR) inhibition for patients with metastatic renal cell carcinoma (mRCC). We measured Ang2 expression in human tissue and plasma, and tested the effect of dual Ang1/2 (trebananib; AMG386) or Ang2 alone (L1-7) inhibition with VEGFR inhibition on murine RCC growth and blood flow. Ang2 levels were higher in human tumors than normal tissues with RCC ranking highest for Ang2 expression across all tumor types tested. Plasma Ang2 was significantly higher in patients with mRCC compared to controls or patients with stage I disease. Plasma Ang2 decreased with sunitinib treatment and increased at time of disease progression. In the RCC mouse, dual Ang1/2 and Ang2 inhibition improved the activity of sunitinib. Combined Ang1/2 and VEGFR inhibition prevented the resumption of blood flow associated with sunitinib resistance. Thus, Ang2 inhibition, independent of Ang1 inhibition, improves the activity of sunitinib and plasma Ang2 increases in the setting of progression on sunitinib possibly contributing to resistance. Further, arterial spin-labeled perfusion magnetic resonance imaging might be a non-invasive marker of the antiangiogenic activity of Ang inhibitors. 相似文献