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Biomedical subjects

J Velek

Publications and source records attributed to J Velek.

At least 19 recordsLinked to original sources
PubMed2006

Peripheral blood mononuclear cell responses to heat shock proteins and their derived synthetic peptides in juvenile idiopathic arthritis patients.

BACKGROUND: Sequence homology and cross reactivity between microbial and human heat shock proteins (hsps) led to the concept that hsps might be involved in the etiopathogenesis of autoimmune diseases. OBJECTIVE: In our study we stimulated peripheral blood mononuclear cells (PBMC) of patients with juvenile idiopathic arthritis (JIA) and healthy controls with various hsp-derived peptides together with the whole molecules of corresponding hsp. METHODS: PBMC were cultured with recombinant human hsp60 (rh-hsp60), rh-hsp70, Mycobacterium bovis hsp65 (M.bovis hsp65), P562-571 human hsp60, P180-188 M. bovis hsp65, P450-463 human hsp70 and P545-554 cytokeratin derived synthetic peptides. Cell responses were measured after incorporation of (3)H-thymidine and expressed as stimulation indices. RESULTS AND CONCLUSION: We found elevated proliferative response to rh-hsp60, M. bovis hsp65 and P562-571 human hsp60 derived peptide in patients with JIA polyarthritis. Significantly elevated proliferation to P180-188 M. bovis hsp65 was found in JIA lasting more than 2 years. None of the particular clinical characteristics (RF, ANA, HLA B27 and disease activity) seemed to be associated with hsp or hsp-derived synthetic peptide proliferative response in the JIA cohort.

Adolescent↗
PubMed2004

Immunogenicity of dendritic cell-based HPV16 E6/E7 peptide vaccines: CTL activation and protective effects.

We have investigated the capacity of cellular vaccines based on dendritic cells loaded with human HPV16 E6/E7 oncoprotein-derived peptides to induce immune responses in vitro and to elicit protective immunity in a murine experimental model mimicking human HPV16-associated carcinomas. Dendritic cells loaded with the HPV16 E6/E7 peptides exhibiting CTL or Th epitopes (E6(41-50), E6(81-90), E6(98-107), E6(130-137), E7(49-57), and E7(44-62)) were able to stimulate in vitro DNA synthesis in syngeneic H-2b spleen cells. The priming capacity of peptide-loaded BMDC and peptide-loaded dendritic cell lines DC2.4 and JAWS II was compared. It has been found that both peptide-loaded BMDC and established dendritic cell lines can activate the syngeneic responder cells, but the priming capacity of BMDC was substantially higher. In the second set of experiments, we have examined the cytolytic activity of syngeneic spleen cells after repeated activation in vitro with BMDC loaded with HPV16 synthetic peptides containing CTL epitopes. Significant cytotoxic responses against HPV16 E6/E7 antigen-expressing TC-1 targets have been found after repeated in vitro activation with all peptides containing the CTL epitopes. In contrast, peptide E7(44-62) harbouring both Th and CTL epitopes induced significant cytotoxic responses already after single in vitro activation. This cytotoxic effect could be enhanced with admixture of the E7(49-57) peptide. Experiments with MHC class I proficient (TC-1, MK16-IFNgamma) and deficient (MK16) target cells revealed that the dendritic cells loaded with the E6/E7 HPV16 peptides activated CTLs in vitro, but not the other cytolytic effector mechanisms. The effectiveness of the peptide-loaded BMDC-based cellular vaccines was also investigated in vivo. C57BL/6 (H-2b) mice were immunized with various peptide-loaded BMDC and subsequently challenged with TC-1 cells. The strongest protective effect was achieved with the BMDC loaded with the peptide E7(44-62) harbouring both CTL and Th epitopes. Mice immunized with the E7(44-62) peptide remained tumour-free after s.c. transplantation of the TC-1 cells and exhibited long-lasting protective immunity, whereas the mice immunized with E6(81-90) and E7(49-57) peptides did not remain tumour-free and exhibited only partial inhibition of tumour growth detectable as depression of the tumour growth curves.

Animals↗
PubMed2004

LVV-hemorphin-7 lowers blood pressure in spontaneously hypertensive rats: radiotelemetry study.

Cardiovascular effects of LVV-hemorphin-7, a member of the family of fragments from beta-chain of human or bovine hemoglobin, were studied in conscious spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats by radiotelemetry. Intraperitoneal injection of hemorphin in a dose of 100 microg/kg significantly decreased blood pressure in SHR, whereas negligible effect was seen in normotensive WKY rats. Blood pressure changes were accompanied by reduction of heart rate. In conclusion, a direct effect of LVV-hemorphin-7 on blood pressure was demonstrated in SHR. These biologically active peptides could be involved in blood pressure regulation especially in hypertensive rats, but the precise mechanism should be elucidated.

Animals↗
PubMed2001

Human and murine humoral immune recognition of multiple peptides from Schistosoma mansoni glyceraldehyde 3-P dehydrogenase is associated with resistance to Schistosomiasis.

Schistosoma mansoni glyceraldehyde 3-phosphate dehydrogenase (SG3PDH) is a target of cellular and humoral immune responses of Brazilian and Egyptian subjects putatively resistant to reinfection with S. mansoni. In an aim to develop a safe, stable and effective vaccine based on this promising molecule, six peptides derived from its primary sequence were selected based on the lowest homology to human G3PDH. The synthetic peptides were tested by ELISA against plasma of humans putatively susceptible or resistant to reinfection with S. mansoni or S. haematobium following chemotherapeutic cure of previous infection. Repeat experiments indicated that the six peptides bear human B-cell epitopes that bind immunoglobulin (Ig)M, IgG1 and IgG3 antibodies. Resistance to reinfection appeared to be significantly associated with humoral immune responses to multiple peptides. This contention was supported by studies in the murine model, whereby we examined the B cell immune responses of Swiss and inbred BALB/c and C57BL/6 mice immunized with recombinant SG3PDH (rSG3PDH) to the six SG3PDH-derived peptides. The serum antibodies of rSG3PDH-immunized Swiss mice were directed to only one of the six peptides tested by ELISA. Antibodies from rSG3PDH-immunized C57BL/6 and BALB/c mice bound, respectively, to four and six out of six peptides. In contrast to Swiss mice, immunization of C57BL/6 and BALB/c mice with rSG3PDH induced protection against challenge cercariae which reached the level of significance (P < 0.05) for BALB/c mice. The data together indicate that host recognition of multiple peptides of a candidate vaccine antigen is necessary for the expression of its ability to contribute to protective immunity against Schistosomiasis.

Adult↗
PubMed2000

Human tumour-associated cell adhesion protein MN/CA IX: identification of M75 epitope and of the region mediating cell adhesion.

MN/CA IX is a cell surface protein, strongly associated with several types of human carcinomas. It exerts activity of carbonic anhydrase and capacity of binding to cell surface receptors. In the present work, we used affinity purified MN/CA IX protein to demonstrate that the cells adhere to immobilized MN/CA IX and that the monoclonal antibody M75 abrogates cell attachment to MN/CA IX. Using synthetic oligopeptides, we identified M75 epitope and located it in the proteoglycan domain, which contains a sixfold tandem repeat of six amino acids GEEDLP. From phage display library of random heptapeptides we identified and chemically synthesized those which compete for the epitope with M75 and inhibit adhesion of cells to MN/CA IX. These heptapeptides might serve as lead compounds for drug design.

Amino Acid Sequence↗
PubMed2000

Somatotropin has no effect on the quantity of guanine nucleotide binding proteins Gqalpha/G11alpha in goat adipose tissue in vivo.

The decapeptide QLNLKEYNLV corresponding to the C-terminus of Gq/G11alpha guanine nucleotide-binding proteins (G-proteins) was synthesized by the solid-phase method and conjugated to keyhole limpet hemocyanin. The rabbits were immunized with these conjugates and an antiserum that reacted specifically with the alpha subunit of Gq/G11 proteins was used in this study. The antiserum exhibited no cross-reactivity with the alpha subunits of stimulatory (Gs) or inhibitory (Gi) G-proteins associated with adenylate cyclase. Immunoblots with the antiserum showed that it specifically recognized the Gq/G11 alpha-proteins in cholate extracts of adipose tissue membranes of goats. Treatment of young castrated male goats with bST had no effect on the quantity of Gq/G11 alpha subunits in adipose tissue and the results thus obtained did not support the idea that the bST signal in adipose tissue is transmitted via Gq/G11 alpha-proteins.

Adipose Tissue↗
PubMed1999

Influence of active immunization against angiotensin AT1 or AT2 receptor on hypertension development in young and adult SHR.

The influence of long-lasting blockade of angiotensin AT1 or AT2 receptors by antibody against the particular receptor peptides on blood pressure and relative heart and kidney weight was studied in spontaneously hypertensive rats (SHR). Young and adult SHR were repeatedly immunized against the sequence 14-23 of angiotensin AT1 receptor from the age of either 1 or 3 months. Other groups of young and adult SHR were immunized against the sequences 37-43 and 106-116 of angiotensin AT2 receptor. Synthetic peptides conjugated to bovine gamma globulin were used as antigens. After 5 months of immunization, blood pressure was measured by the direct method. All immunized animals produced antibodies against the particular peptides. At the end of immunization, the blood pressure was significantly decreased in SHR immunized in youth against angiotensin AT1 receptor peptide, although no difference in heart and kidney hypertrophy was observed compared to sham-immunized SHR. The immunization against angiotensin AT1 receptor peptide in adulthood as well as the immunization against angiotensin AT2 receptor peptides in youth or in adulthood affected neither blood pressure nor heart and kidney weight. No influence of immunization on the studied parameters was observed in normotensive WKY rats. Angiotensin AT1 receptors play a more important role in the pathogenesis of spontaneous hypertension than angiotensin AT2 receptors. The blockade of angiotensin AT1 receptors by active immunization against the receptor peptide attenuated hypertension development in young SHR but did not modify the already established hypertension in adult SHR.

Aging↗
PubMed1996

Blockade of AT1 receptors by specific antibody attenuated hypertension development in young spontaneously hypertensive rats.

The influence of chronic angiotensin AT1 receptor blockade by specific antibody on the development of genetic hypertension was studied in young spontaneously hypertensive rats (SHR). The immunization of 4-week-old SHR with a small part of the angiotensin AT1 receptor molecule attenuated the development of hypertension in these animals. After five subcutaneous injections of the antigen both systolic and diastolic blood pressures were significantly lower (p < 0.005) in immunized SHR compared to sham-immunized SHR. No effect on blood pressure was seen in immunized Wistar-Kyoto control rats. We conclude that renin-angiotensin system might be partially involved in the development of hypertension in young spontaneously hypertensive rats because it can be attenuated by a specific antibody raised against a part of the angiotensin AT1 receptor.

Angiotensin II↗
PubMed1995

[Hormonal treatment of non-malignant diseases of the breast].

The authors summarize possibilities of hormonal treatment of non-malignant diseases of the mammary gland. Attention is paid in particular to progestins (progesterone, derivatives of hydroxyprogesterone, derivatives of 19 nor-testosterone), their combination with estrogens (hormonal contraceptives, minipills) and hormonal substitution therapy, methods of chemoprevention of mammary cancer. In conjunction with the subject the authors mention also the possibility to use some other preparations (Danazol, Gonadotropin releasing hormones, tamoxifen etc.). The authors submit their own pattern of hormonal treatment of non-malignant diseases of the mammary gland.

Breast Diseases↗
PubMed1994

Angiotensin AT1 receptor blockade by specific antibody prevented two-kidney, one-clip renal hypertension in the rat.

The effect of chronic angiotensin AT1 receptor blockade by a specific antibody on the development of two-kidney, one-clip renal hypertension was studied in Wistar rats. Renal artery constriction resulted in a fast and large increase in blood pressure in comparison with that of control rats. On the other hand, the pre-immunization of rats with a small part of the angiotensin AT1 receptor completely prevented the development of renal hypertension. We conclude that the development of two-kidney, one-clip renal hypertension can be blocked by a specific antibody raised against a part of the angiotensin AT1 receptor.

Amino Acid Sequence↗
PubMed1994

Semisynthetic insulin analogues modified in positions B24, B25 and B29.

New semisynthetic analogues of human insulin, modified in the C-terminal region of the B-chain, were prepared to refine our understanding of the importance of particular amino acid residues in the expression of hormone biological properties. The following insulin analogues were synthesized by trypsin-catalyzed peptide-bond formation between the C-terminal arginineB22 of des-octapeptide(B23-B30)-insulin and synthetic octapeptides with the epsilon-amino group of lysineB29 protected by a phenylacetyl group: [L-Lys(Pac)B29]insulin, [D-PheB24,B25,L-Lys(Pac)B29]insulin and [D-Phe(p-Et)B24, L-Lys(Pac)B29]insulin. Enzymatic deprotection using immobilized penicillin amidohydrolase yielded: human insulin, [D-PheB24,B25]insulin and [DPhe(p-Et)B24]-insulin. Biological in vitro potencies (specific binding to cultured human lymphocytes IM-9 and lipogenic potency in isolated rat adipocytes) of the semisynthetic analogues were estimated, ranging from 0.2 to 100% relative to porcine insulin.

Adipocytes↗
PubMed1991

Specificity studies on retroviral proteinase from myeloblastosis-associated virus.

The specificity of the p15 proteinase of myeloblastosis-associated virus (MAV) was tested with nonviral high molecular weight substrates and with synthetic peptides. Peptides with sequences spanning known cleavage sites in viral polyproteins of Rous sarcoma virus (RSV) and avian leukemia viruses, as well as in BSA and HSA, were synthesized, and the rate of their cleavage by the MAV proteinase was compared. Synthetic peptides require for successful cleavage at least 4 residues at the N-terminal side and 3 residues at the C-terminal side. The proteinase shows a preference for hydrophobic residues with bulky side chains (Met, Tyr, Phe) in P3, although Arg and Gln can also be accepted. Small hydrophobic residues are required in P2 and P2', and large hydrophobic residues (Tyr, Met, Phe/p-nitro-Phe) are preferred in both P1 and P1'. The difference between the specificity of the p15 proteinase and that of the HIV-1 proteinase mostly pertains to position P2' of the substrate, where bulkier side chains are accepted by the HIV-1 proteinase (Richards et al., 1990). A good chromogenic substrate for the MAV and RSV proteinases was developed and used to further characterize the MAV proteinase activity with respect to ionic strength and pH. The activity of the proteinase is strongly dependent on ionic strength and pH. Both the kcat and Km values contribute to a higher cleavage efficiency at higher salt concentrations and show a bell-shaped pH dependence curve with a sharp maximum at pH 5.5 (kcat) and 6.5 (Km).

Amino Acid Sequence↗