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The TLR7 agonists imiquimod and gardiquimod improve DC-based immunotherapy for melanoma in mice

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cellular & Molecular Immunology 2010年第5期7卷 381-388页

作者： Fang Ma Jianhua Zhang Jian Zhang Cai Zhang Institute of Immunopharmacology and immunotherapy School of Pharmaceutical SciencesShandong UniversityJinanChina

Toll-like receptors(TLRs)are a family of highly conserved germline-encoded pattern-recognition receptors that are essential for host immune *** ligands represent a promising class of immunotherapeutics or vaccine adjuvants with the potential to generate an effective antitumor immune *** TLR7/8 agonists have aroused interest because they not only activate antigen-presenting cells but also promote activation of T and natural killer(NK)***,the exact mechanism by which stimulation of these TLRs promotes immune responses remains unclear,and different TLR7/8 agonists have been found to induce different *** this study,we demonstrate that both gardiquimod and imiquimod promote the proliferation of murine splenocytes,stimulate the activation of splenic T,NK and natural killer T(NKT)cells,increase the cytolytic activity of splenocytes against B16 and MCA-38 tumor cell lines,and enhance the expression of costimulatory molecules and IL-12 by macrophages and bone marrow-derived dendritic cells(DCs).In a murine model,both agonists improved the antitumor effects of tumor lysate-loaded DCs,resulting in delayed growth of subcutaneous B16 melanoma tumors and suppression of pulmonary ***,we found that gardiquimod demonstrated more potent antitumor activity than *** results suggest that TLR7/8 agonists may serve as potent innate and adaptive immune response modifiers in tumor *** importantly,they can be used as vaccine adjuvants to potentiate the efficiency of DC-based tumor immunotherapy.

关键词： antitumor therapy DC vaccine melanoma TLR TLR7/8 agonist

High efficiency production and genomic in situ hybridization analysis of Brassica aneuploids and homozygous plants

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Science China(Life Sciences) 2003年第1期46卷 104-112页

作者：李再云 M.Ceccarelli S.Minelli A.Contento 刘焰 P.G.Cionini National Key lab of Crop Genetic Improvement Department of AgronomyHuazhong Agricultural University Department of cellular and Molecular Biology University of Perugia

Interspecific and intergeneric hybridizations have been widely used in plant genetics and breeding to construct stocks for genetic analysis and to introduce into crops the desirable traits and genes from their relatives. The intergeneric crosses between Brassica juncea (L.) Czern. & Coss., B. carinata A. Braun and Orychophragmus violaceus (L.) O. E. Schulz were made and the plants produced were subjected to genomic in situ hybridization analysis. The mixoploids from the cross with B. juncea were divided into three groups. The partially fertile mixoploids in the first group (2n = 36—42) mainly contained the somatic cells and pollen mother cells (PMCs) with the 36 chromosomes of B. juncea and additional chromosomes of O. violaceus. The mixoploids (2n = 30—36) in the second and third groups were morphologically quite similar to the mother plants B. juncea and showed nearly normal fertility. The plants in the second group produced the majority of PMCs (2n = 36) with their chromosomes paired and segregated normally, but 1—4 pairs of the O. violaceus chromosomes were included in some PMCs. The plants in the third group produced only PMCs with the 36 B. juncea chromosomes, which were paired and segregated normally. The mixoploids (2n = 29—34) from the cross with B. carinata produced the majority of PMCs (2n = 34) with normal chromosome pairing and segregation, but some plants had some PMCs with 1—3 pairs of chromosomes from O. violaceus and other plants had only PMCs with the B. carinata chromosomes. The Brassica homozygous plants and aneuploids with complete or partial chromo-some complements of Brassica parents and various numbers of O. violaceus chromosomes were derived from these progeny plants. The results in this study provided the molecular cytogenetic evidence for the separation of parental genomes which was previously proposed to occur in the hybridizations of these two genera.

关键词： Brassica species, Orychophragmus violaceus, aneuploids, intergeneric hybrids, genome separation.

Mast cells as rapid innate sensors of cytomegalovirus by TLR3/TRIF signaling-dependent and -independent mechanisms

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cellular & Molecular Immunology 2015年第2期12卷 192-201页

作者： Marc Becker Niels AW Lemmermann Stefan Ebert Pamela Baars Angelique Renzaho Jurgen Podlech Michael Stassen Matthias J Reddehase Institute for Immunology Research Center for Immunotherapy (FZI) at the University Medical Center of the Johannes Gutenberg University Mainz Germany Institute for Virology Research Center for Immunotherapy (FZI) at the University Medical Center of the Johannes Gutenberg University Mainz Germany These authors contributed equally to this work Co-senior authors

The succinct metaphor,‘the immune system＇s loaded gun＇, has been used to describe the role of mast cells （MCs） due to their storage of a wide range of potent pro-inflammatory and antimicrobial mediators in secretory granules that can be released almost instantly on demand to fight invaders. Located at host-environment boundaries and equipped with an arsenal of pattern recognition receptors, MCs are destined to be rapid innate sensors of pathogens penetrating endothelial and epithelial surfaces. Although the importance of MCs in antimicrobial and antiparasitic defense has long been appreciated, their role in raising the alarm against viral infections has been noted only recently. Work on cytomegalovirus （CMV） infection in the murine model has revealed MCs as players in a novel cross-talk axis between innate and adaptive immune surveillance of CMV, in that infection of MCs, which is associated with MC degranulation and release of the chemokine CCL5, enhances the recruitment of protective CD8 T cells to extravascular sites of virus replication, specifically to lung interstitium and alveolar epithelium. Here, we have expanded on these studies by investigating the conditions for MC activation and the consequent degranulation in response to host infection. Surprisingly, the data revealed two temporally and mechanistically distinct waves of MC activation： an almost instant indirect activation that depended on TLR3/TRIF signaling and delayed activation by direct infection of MCs that did not involve TLR3/rRIF signaling. Cell type-specific Cre-recombination that yielded eGFP-expressing reporter virus selectively originating from MCs identified MC as a new in vivo, first-hit target cell of productive murine CMV infection.

关键词： cytomegalovirus innate immunity mast cells mast cell-specific Cre-recombination TLR3FrRIF-signaling

Is there a close relationship between synonymous codon bias and codon-anticodon binding strength in human genes?

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Chinese Science Bulletin 2001年第12期46卷 1015-1019页

作者： Xiufan Shi Jingfei Huang Chongrong Liang Shuqun Liu Jun Xie Ciquan Liu Chinese Acad Sci Kunming Inst Zool Cellular & Mol Evolutionary Lab Kunming 650223 Peoples R China

Synonymous codon bias has been examined in 78 human genes (19967 codons) and measured by relative synonymous codon usage (RSCU). Relative frequencies of all kinds of dinucleotides in 2,3 or 3,4 codon positions have been calculated, and codon-anticodon binding strength has been estimated by the stacking energies of codon-anticodon bases in Watson-Crick pairs. The data show common features in synonymous codon bias for all codon families in human genes: all C-ending codons, which possess the strongest codon-anticodon binding energies, are the most favored codons in almost all codon families, and those codons with medium codon-anticodon binding energies are avoided. Data analysis suggests that besides isochore and genome signature, codon-anticodon binding strength may be closely related to synonymous codon choice in human genes. The join-effect of these factors on human genes results in the common features in codon bias.

关键词： codon bias synonymous codons human genes.

PD-1^(+)and TIM-3^(+)T cells widely express commonγ-chain cytokine receptors in multiple myeloma patients,and IL-2,IL-7,IL-15 stimulation up-regulates PD-1 and TIM-3 on T cells

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Oncology Research 2024年第10期32卷 1575-1587页

作者： EGOR V.BATOROV ALISA D.INESHINA TATIANA A.ARISTOVA VERA V.DENISOVA SVETLANA A.SIZIKOVA DARIA S.BATOROVA GALINA Y.USHAKOVA EKATERINA Y.SHEVELA ELENA R.CHERNYKH laboratory of cellular immunotherapy Research Institute of Fundamental and Clinical ImmunologyNovosibirsk630099Russia V.Zelman Institute of Medicine and Psychology Novosibirsk National Research State UniversityNovosibirsk630090Russia Department of Hematology and Bone Marrow Transplantation Research Institute of Fundamental and Clinical ImmunologyNovosibirsk630099Russia

Background:Immune checkpoint ligand-receptor interactions appear to be associated with multiple myeloma(MM)***,previous studies showed the possibility of PD-1 and TIM-3 expression on T cells upon stimulation with commonγ-chain family cytokines in vitro and during homeostatic *** aim of the present work was to study the impact of homeostatic proliferation on the expansion of certain T cell subsets upregulating PD-1 and TIM-3 checkpoint ***:The expression of CD25,CD122,CD127 commonγ-chain cytokine receptors,phosphorylated signal transducer and activator of transcription-5(pSTAT5)and eomesodermin(EOMES)was comparatively assessed with flow cytometry in PD-1-and TIM-3-negative and positive T cells before the conditioning and during the first post-transplant month in peripheral blood samples of MM ***:Substantial proportions of PD-1-and TIM-3-positive T lymphocytes expressed commonγ-chain cytokine receptors and *** of cytokine receptor expressing cells were significantly higher within TIM-3+T cells compared to PD-1+TIM-3−*** proportions of both PD-1-/TIM-3-negative and positive CD8+T cells express EOMES,while only moderate frequencies of CD4+PD-1+/TIM-3+T cells up-regulate this transcription ***,the surface presence of CD25 and intranuclear expression of EOMES in CD4+T cells were mutually exclusive regardless of PD-1 and TIM-3 *** stimulation with commonγ-chain cytokines up-regulates PD-1 and TIM-3 during the proliferation of initially PD-1/TIM-3-negative T cells but fails to expand initially PD-1+and TIM-3+T cell subsets in ***:Both PD-1 and TIM-3 expressing T cells appear to be able to respond to homeostatic cytokine *** in commonγ-chain cytokine receptor expression between PD-1+and TIM-3+T cells may reflect functional dissimilarity of these cell *** blockade appears to alleviate lymphopenia-induced prolif

关键词： Autologous hematopoietic stem cell transplantation(AHSCT) CD25 CD122 Eomesodermin(EOMES) Homeostatic proliferation

Identification of protein superfamily from structure-based sequence motif

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Chinese Science Bulletin 2002年第16期47卷 1377-1381页

作者： Jingfei Huang Ciquan Liu Chinese Acad Sci Kunming Inst Zool Key Lab Cellular & Mol Evolut Kunming 650223 Peoples R China

The structure-based sequence motif of the distant proteins in evolution, protein tyrosine phosphatases (PTP) I and Ⅱ superfamilies, as an example, has been defined by the structural comparison, structure-based sequence alignment and analyses on substitution patterns of residues in common sequence conserved regions. And the phosphatases I and Ⅱ can be correctly identified together by the structure-based PTP sequence motif from SWISS-PROT and TrEBML databases. The results show that the correct rates of identification are over 98%. This is the first time to identify PTP I and Ⅱ together by this motif.

关键词： protein tyrosine phosphatases sequence motif sequence identification of superfamily hypothetical proteins molecular evolu-tion.

Reactivity against microsatellite instability-induced frameshift mutations in patients with inflammatory bowel disease

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World Journal of Gastroenterology 2015年第1期21卷 221-228页

作者： Florian Kuehn Ernst Klar Anja Bliemeister Michael Linnebacher Department of General ThoracicVascular and Transplantation SurgeryMolecular Oncology and ImmunotherapyUniversity of Rostock18057 RostockGermany

AIM: To analyze the cellular immune response towards microsatellite-instability(MSI)-induced frameshift-peptides(FSPs) in patients suffering from inflammatory bowel disease(IBD) with and without thiopurine-based immunosuppressive ***: Frequencies of peripheral blood T cell responses of IBD patients(n = 75) against FSPs derived from 14 microsatellite-containing candidate genes were quantified by interferon-γ enzyme-linked immunospot.T cells derived from 20 healthy individuals served as ***: Significant T cell reactivities against MSIinduced FSPs were observed in 59 of 75 IBD patients(78.7%).This was significantly more as we could observe in 20 healthy controls(P = 0.001).Overall,the reactivity was significantly influenced by thiopurine treatment(P = 0.032) and duration of disease(P = 0.002) but not by duration or cumulative amount of thiopurine therapy(P = 0.476).Unexpected,15 of 24(62.5%) IBD patients without prior thiopurine treatment also showed increased FSP-specific immune responses(P = 0.001).CONCLUSION: These findings propose FSPs as potential novel class of inflammation-associated antigens and this in turn may have implications for screening,diagnosis as well as clinical management of patients suffering from IBD and other inflammatory conditions.

关键词： Microsatellite-instability Inflammatory bowel dise

Recombinant Expression of a Novel Human Transcriptional Repressor HMBOX1 and Preparation of Anti-HMBOX1 Monoclonal Antibody

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cellular & Molecular Immunology 2009年第4期6卷 261-268页

作者： Jun Dai Longyan Wu Cai Zhang Xiaodong Zheng Zhigang Tian Jian Zhang Institute of Immunopharmacology ＆ immunotherapy School ofPharmaceutical Sciences Shandong University Jinan 250012 China Institute of Immunology School of Life Sciences University of Science ＆Technology of China Hefei 230027 China Institute of Immunopharmacology ＆ immunotherapy School of Pharmaceutical Sciences Shandong University 44 Wenhua West Road Jinan 250012 China

HMBOX1 was a novel transcription factor possibly involving in function of pancreas and cytotoxicity of NK cells. For function determination, recombinant human HMBOX1 protein was obtained and purified, and the monoclonal antibodies against HMBOX1 were prepared. The full-length cDNA fragment encoding HMBOX1 was amplified from NK-92 cells and inserted into prokaryotic expression vector pET22b. The pET22b-HMBOX1-6his vector was then transformed into E. coli Rosetta （DE3） and induced by 1 mM IPTG for 4 h at 37℃. The fusion HMBOX1 protein was mainly expressed in inclusion bodies, which was purified and refolded using Ni^2＋-affinity chromatography. With the purified fusion HMBOX1 protein as antigen, monoclonal antibodies against HMBOX1 were generated, providing a potentially useful tool for further study in HMBOX1 functions. Using these anti-HMBOX1 mAbs, we identified that HMBOX1 is located in both cytoplasm and nucleus and could be detected in 10 human normal tissues, including cerebrum, pancreas, kidney and liver tissues. Moreover, the expression in hepatic carcinoma was significantly lower than that in adjacent tissues.

关键词： HMBOX1 expression purification renaturation monoclonal antibody

Engineering bionic T cells: signal 1, signal 2, signal 3, reprogramming and the removal of inhibitory mechanisms

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cellular & Molecular Immunology 2020年第6期17卷 576-586页

作者： Iñaki Etxeberria Irene Olivera Elixabet Bolaños Asunta Cirella Álvaro Teijeira Pedro Berraondo Ignacio Melero Program of Immunology and immunotherapy Center for Applied Medical Research(CIMA)PamplonaSpain Navarra Institute for Health Research(IDISNA) PamplonaSpain Centro de Investigación Biomédica en Red de Cáncer(CIBERONC) MadridSpain Department of Immunology and immunotherapy Clínica Universidad de NavarraPamplonaSpain

Gene engineering and combinatorial approaches with other cancer immunotherapy agents may confer capabilities enabling full tumor rejection by adoptive T cell therapy(ACT).The provision of proper costimulatory receptor activity and cytokine stimuli,along with the repression of inhibitory mechanisms,will conceivably make the most of these treatment *** this sense,T cells can be genetically manipulated to become refractory to suppressive mechanisms and exhaustion,last longer and differentiate into memory T cells while endowed with the ability to traffic to malignant *** antitumor effects can be dramatically augmented with permanent or transient gene transfer maneuvers to express or delete/repress genes.A combination of such interventions seeks the creation of the ultimate bionic T cell,perfected to seek and destroy cancer cells upon systemic or local intratumor delivery.

关键词： Adoptive cell therapy T cell engineering Cancer immunotherapy