Staff. A.Yu. Sazykin, PhD, S.V. Khlgatian, PhD (part-time), D.V. Kuprash, PhD (part-time), Yu.V. Shebzukhov, PhD (till 2005), I.N. Lavrik, PhD (till 2006), M.A. Goncharskaya, PhD (part-time), R.Sh. Kazarian, T.S. Kadachigova, G.I Efremov, P.V. Belousov (part-time), L.N. Drutskaya, A.A. Kruglov, A.R. Galimov.

Main lines of investigation. Serologic identification and characterization of novel human tumor antigens. Physiological functions of TNF family cytokines (TNF, LTa, LTb) and the investigation of consequences of TNF blocking in vivo and development of chymeric antibodies for therapeutical blocking of TNF in case of its overexpression in autoimmune and infectious diseases. Improvement of genetic knock-out technology.  Engineering mice with mutant cytochrome C gene.
Participation in research projects (programs) and grant support. The project on identification and characterization of tumor antigens was carried out in close collaboration with Ludwig Institute for Cancer Research, New York, NY, USA; N.N. Blokhin Cancer Research Center, Moscow;  Engelhardt Institute of Molecular Biology, Moscow; Medizinische Klinik, Haematologie-Onkologie, Hospital Nordwest, Frankfurt, Germany. The project on generation of TNF- and lymphotoxin-deficient mice was carried out in collaboration with National Cancer Institute, Frederick, MD, USA and Engelhardt Institute of Molecular Biology, Moscow, Russia. The Department is involved in the organization  "Oncoimmunology" educational program (www.oncoimmunology.ru) sponsored by Cancer Research Institute (New York). Research grants: Ludwig Institute for Cancer Research (LICR), Russian Foundation of Basic Research (RFBR), Federal Agency for Science and Innovations (FASI).

Major research advances. Serologic screening of expression cDNA libraries from renal cancer,  breast cancer, ovary cancer, colorectal cancer cell lines and normal testis (approximately 2000000 clones tested, out of which 443 serologically positive clones were submitted to Cancer Immunome Database, SEREX project) Three novel human cancer antigens were discovered, and seven previously described proteins were shown to be putative cancer antigens [Lagarkova et al., 2000; Kuimov et al., 2001; Shebzukhov et al., 2005a, Shebzukhov et al., 2005b]. We developed an improved method to detect serological responses to tumor-associated antigens (SMARTA) using miniarrays [Lagarkova et al., 2003]. A panel of cancer-specific antigens suitable for serologic diagnostics and monitoring of melanoma, renal cancer, breast cancer, ovary cancer, lung cancer and colon cancer has been established. A sizable collection of clinical specimens  (blood serum samples and tissue specimens from patients with different cancers) has been  characterized for reactivity against  the panel of cancer-specific antigens.

A unique panel of mice with complete and tissue-specific inactivation of tumor necrosis factor (TNF) and lymphotoxin (LT) genes was generated. In particular, physiologic functions of LT produced by B-lymphocytes [Tumanov et al., 2002] and of TNF produced by macrophages and T-cells [Grivennikov et al., 2005], as well as cooperation of TNF and LT in vivo [Kuprash, et al., 2002] have been established. Molecular mechanism of immunosuppressant cyclosporin A influence on expression of LT genes was investigated [Kuprash, et al., 2002]. Mouse monoclonal antibodies against TNF were raised and sequenced [Radko et al., Russ. J. Immunol. 7: 371, 2002], and they are being engineered to us as a TNF inhibitor in vivo. A novel animal model to study the consequences of TNF blockade vivo has been generated: mice “humanized” for  TNF/LT locus. These mice produce human TNF which completely compensates for the loss of its mouse counterpart in immune and host defense functions, lymphoid organs architecture and in models of TNF-mediated systemic toxicity.

Scientific recognition. Perigrinus Award from Berlin-Brandenburg Academy of Sciences (S.A. Nedospasov), Helmholtz-Humboldt Research Award (S.A. Nedospasov), Russian Biochemical Society for Young Scientists Award and European Academy Award (D.V. Kuprash).

List of publications:

1. Regulation of cytokine genes transcription:

1.    Stuber F, Udalova IA, Book M, Drutskaya LN, Kuprash DV, Turetskaya RL, Schade FU, Nedospasov SA. -308 tumor necrosis factor (TNF) polymorphism is not associated with survival in severe sepsis and is unrelated to lipopolysaccharide inducibility of the human TNF promoter. J.Inflamm. 1995; 46: 42-50.
2.    Kuprash DV, Osipovich OA, Pokholok DK, Alimzhanov MB, Biragyn A, Turetskaya RL, Nedospasov SA. Functional analysis of the lymphotoxin-beta promoter. Sequence requirements for PMA activation. J.Immunol. 1996; 156: 2465-2472.
3.    Baer M, Dillner A, Schwartz RC, Sedon C, Nedospasov SA, Johnston PF. TNF alpha transcription in macrophages is attenuated by an autocrine factor that preferentially induces NF-kB p50. Mol.Cell.Biol. 1998; 18:5678-5689.
4.    Kaijzel EL, van Krugten MV, Brinkman BM, et al. Functional analysis of a human tumor necrosis factor alpha (TNF-alpha) promoter polymorphism related to joint damage in rheumatoid arthritis. Mol Med 1998; 4:724-733.
5.    Udalova IA, Knight JC, Vidal V, Nedospasov SA, Kwiatkowski D. Complex NF-kappaB interactions at the distal tumor necrosis factor promoter region in human monocytes. J.Biol.Chem. 1998; 273: 21178-21186.
6.    Kuprash DV, Udalova IA, Turetskaya RL, Kwiatkowski D, Rice NR, Nedospasov SA. Similarities and differences between human and murine TNF promoters in their response to lipopolysaccharide. J Immunol 1999; 162: 4045-4052.
7.    Kuprash DV, Boitchenko VE, Yarovinsky FO, Rice NR, Nordheim A, Ruhlmann A, Nedospasov SA. Cyclosporin A blocks the expression of lymphotoxin alpha, but not lymphotoxin beta, in human peripheral blood mononuclear cells. Blood. 2002; 100(5): 1721-1727.

2. Physiologic functions of cytokines (in genetically knock-out mice):

8.    Alimzhanov MB, Kuprash DV, Kosco-Vilbois MH, Luz A, Turetskaya RL, Tarakhovsky A, Rajewsky K, Nedospasov SA, Pfeffer K. Abnormal development of secondary lymphoid tissues in lymphotoxin beta- deficient mice. Proc Natl Acad Sci U S A 1997; 94:9302-9307.
9.    Endres R, Alimzhanov MB, Plitz T, et al. Mature follicular dendritic cell networks depend on expression of lymphotoxin beta receptor by radioresistant stromal cells and of lymphotoxin beta and tumor necrosis factor by B cells. J Exp Med 1999; 189:159-168.
10.    Ito D, Back TC, Shakhov AN, Wiltrout RH, Nedospasov SA. Mice with a Targeted Mutation in Lymphotoxin-alpha Exhibit Enhanced Tumor Growth and Metastasis: Impaired NK Cell Development and Recruitment. J Immunol 1999; 163: 2809-2815.
11.    Kuprash DV, Alimzhanov MB, Tumanov A, Anderson AO, Pfeffer K, Nedospasov SA. TNF and Lymphotoxin beta cooperate in the maintenance of secondary lymphoid tissue microarchitecture but not in the development of lymph nodes. J Immunol 1999; 163: 6575-6580.
12.    Drize NI, Drutskaya MS, Gerasimova LP, Manakova TE, Chertkov IL, Turetskaya RL, Kuprash DV, Nedospasov SA. Changes in the hemopoietic system of mice deficient for tumor necrosis factor or lymphotoxin-alpha. Bull Exp Biol Med [Russian] 2000; 130 (7): 76-79.
13.    Shakhov AN, Lyakhov IG, Tumanov AV, et al. Gene profiling approach in the analysis of lymphotoxin and TNF deficiencies. J Leukoc Biol 2000; 68: 151-157.
14.    Shakhov AN, Rubtsov AV, Lyakhov IG, Tumanov A, Nedospasov SA. SPLASH (PLA211D), a novel member of phospholipase A2 family, is associated with lymphotoxin deficiency. Genes Immunity 2000; 1: 191-199.
15.    Drutskaya MS, Kuprash DV, Nedospasov SA, Chertkov IL, Drize NI. Abnormal hemopoiesis in long-term bone marrow culture from tumor necrosis factor-deficient mice. Bull Exp Biol Med [Russian] 2001 Feb; 131(2): 184-187.
16.    Shakhov AN, Nedospasov SA. Expression profiling in knockout mice: lymphotoxin versus tumor necrosis factor in the maintenance of splenic microarchitecture. Cytokine Growth Factor Rev 2001; 12: 107-119.
17.    Hehlgans T, Stoelcker B, Stopfer P, Muller P, Cernaianu G, Guba M, Steinbauer M, Nedospasov SA, Pfeffer K, Mannel DN. Lymphotoxin-beta receptor immune interaction promotes tumor growth by inducing angiogenesis. Cancer Res. 2002 62: 4034-40
18.    Kuprash D.V. Alimzhanov M.B., Tumanov A.V., Ito D., Abe, K., Turetskaya, R.L., Marino, M.W., Anderson A.O., Rajewsky K., Pfeffer K. and Nedospasov S.A. Mutiple deficiencies in immune functions in mice with complete neo-free deletion of the TNF/LT locus (TNF/LTbeta/LTalpha triple-deficient mice). Mol. Cell. Biol., 2002, 22:8626-8634.
19.    Tumanov, A.V., Kuprash, D.V., Lagarkova, M.A., Grivennikov, S.I., Abe, K., Shakhov, A.N., Drutskaya. L.N., Stewart C.L., Chervonsky, A.V. and Nedospasov, S.A. Distinct Role of Surface Lymphotoxin Expressed by B Cells in the Organization of Secondary Lymphoid Tissues. Immunity 2002 17(3): 239-250.
20.    Abe, K., Yarovinsky, F., Murakami, T., Ito, D., Tumanov, A.V., Pfeffer, K., Shakhov, A.N., Kuprash, D.V., Komschlies, K.L., and Nedospasov, S.A. Distinct role of TNF/LT subfamily of cytokines in the development and recruitment of dendritic cells. Blood, 2003, 101:1477-1483.
21.    Grivennikov SI, Tumanov AV, Liepinsh DJ, Kruglov AA, Marakusha BI, Shakhov AN, Murakami T, Drutskaya LN, Forster I, Clausen BE, Tessarollo L, Ryffel B, Kuprash DV, Nedospasov SA. Distinct and non-redundant in vivo functions of TNF produced by T cells and macrophages/neutrophils: protective and deleterious effects. Immunity 2005, 22: 93-104.
22.    Liepinsh DJ, Grivennikov SI, Klarmann KD, Lagarkova MA, Drutskaya MS, Lockett SJ, Tessarollo L, McAuliffe M, Keller JR, Kuprash DV, Nedospasov SA. Novel lymphotoxin alpha (LTalpha) knockout mice with unperturbed tumor necrosis factor expression: reassessing LTalpha biological functions. Mol Cell Biol 2006, 26:4214-4225.

3. Human cancer antigens.

23.    Lagarkova MA, Boitchenko VE, Mescheryakov AA, Kashkarova UA, Nedospasov SA. Human cortactin as putative cancer antigen. Oncogene 2000; 19: 5204-5207.
24.    Kuimov AN, Kuprash DV, Petrov VN, Vdovichenko KK, Scanlan MJ, Jongeneel CV, Lagarkova MA, Nedospasov SA. Cloning and characterization of TNKL, a member of tankyrase gene family. Genes Immun 2001; 2: 52-55.
25.    Koroleva EP, Lagarkova MA, Mesheryakov AA, Scanlan MJ, Old LJ, Nedospasov SA, Kuprash DV. Serological identification of antigens assosiated with renal cell carcinoma. Russian Journal of Immunology, 2002, 7(3):229-238.
26.    Lagarkova MA, Koroleva EP, Kuprash DV, Boitchenko VE, Kashkarova UA, Nedospasov SA, Shebzukhov YV. Evaluation of humoral response to tumor antigens using recombinant expression-based serological mini-arrays (SMARTA). Immunology Letters, 2003, 85(1):71-74.
27.    Koroleva EP, Lagarkova MA, Khlgatian SV, Shebzukhov YV, Mescheryakov AA, Lichinitser MR, Nedospasov SA, Kuprash DV. Serologic analysis of repertoire of human cancer antigens and autoantigens. Mol. biologiya [Russian], 2004, 38: 1-6.
28.    Shebzukhov YV, Koroleva  EP, Khlgatian SV, Belousov PV, Kuz’mina KE, Radko BV, Longpre F, Lagarkova MA, Kadachigova TS, Gurova OV, Mescheryakov AA, Lichinitser MR, Knuth A, Jager E, Kuprash DV, and Nedospasov SA. Humoral immune response to a non-conserved C-terminal part of human histone deacetylase 3 in colon cancer patients.  Int. J. Cancer 2005a; 117(5):800-806.
29.    Shebzukhov YV, Koroleva EP, Khlgatian SV, Lagarkova MA, Meshcheryakov AA, Lichinitser MR, Karbach J, Jager E, Kuprash DV, Nedospasov SA.   Humoral immune response to thymidylate synthase in colon cancer patients after 5-FU chemotherapy. Immunol Lett. 2005b; 100(1): 88-93.

4. Other research in molecular immunology and hematology.

30.    Schroeder HW Jr, Zhu ZB, March RE, Campbell RD, Berney SM, Nedospasov SA, Turetskaya RL, Atkinson TP, Go RC, Cooper MD, Volanakis JE. Susceptibility locus for IgA deficiency and common variable immunodeficiency in the HLA-DR3, -B8, -A1 haplotypes. Mol Med. 1998, 4:72-86.
31.    Korobko VG, Boichenko VE, Kuprash DV, Turetskaia RL, Nedospasov SA. [Heterologous expression of murine lymphotoxins in Escherichia coli and preparation of antibodies]. Bioorg Khim. 1999 Apr; 25(4):270-274.
32.    Boitchenko VE, Korobko VG, Prasolov VS, Kravchenko VV, Kuimov AN, Turetskaya RL, Kuprash DV, Nedospasov SA. Immunodetection of murine lymphotoxins in eukaryotic cells. Russian J Immunol 2000; 5:259-266.
33.    Eskdale J, Turetskaya RL, Armstrong C, Kuprash DV, Nedospasov SA, Gallagher G. A polymorphic microsatellite marker in the human p55 TNF receptor, CD120a. Genes Immun 2000; 1: 228-230.
34.    Б Boitchenko VE, Alimzhanov MB, Turetskaya RL, Ruhlmann A, Nordheim A, Kuprash DV, Nedospasov SA. [Comparative Characterization of Lymphotoxin Transcripts from Human B- and T-Cell Lines, Peripheral Lymphocytes, and Normal Tissues]. Mol Biol (Mosk) 2001; 35: 115-121.
35.    Radko BV, Boitchenko VE, Nedospasov SA, Korobko VG. Characterization of the genes encoding variable light and heavy chains of the high-affinity monoclonal antibody against human tumor necrosis factor. Russian Journal of Immunology, 2002, 7(4): 371-374.

5. Reviews.

36.    Nedospasov SA, Grivennikov SI, Kuprash DV. Physiologic roles of members of the TNF and TNF receptor families as revealed by knockout models. In: "Contemporary immunology: Cytokine Knockouts" 2nd edition, Giamila Fantuzzi, Ed., 2003, Humana Press, Inc., Totowa, NJ, 439-460.
37.    Nedospasov SA, Kuprash DV. [Cytokines, their receptors and intracellular signal transduction]. In: Cancerogenesis [Russian], DG Zaridze ed., 2004, Meditsina, Moscow, 158-168.
38.    Nedospasov SA, Lavrik IN. [Mechanisms of activation of programmed cell death via death receptors]. In: Cancerogenesis [Russian], DG Zaridze ed., 2004, Meditsina, Moscow, 168-179.