Evidence suggests that beta-amyloid (Abeta) has chemokine-like properties and may act through formyl chemotactic receptors (FPR) to induce pathophysiologically important functional changes in Alzheimer's disease (AD) microglia. We have shown that Abeta 1-42, fibrillar Abeta 1-40, and Abeta 25-35 potentiate the release of interleukin-1beta (IL-1beta) from LPS activated human THP-1 monocytes [26] and LPS primed rat microglia. Moreover, Abeta-stimulated IL-1beta secretion seems to be receptor mediated because it is calcium dependent and requires activation of specific G-proteins [27]. Thus, we have evaluated the ability of Abeta 1-42 to mimic formyl chemotactic peptides in stimulating IL-1beta release from THP-1 monocytes. Several of the formyl chemotactic peptides and Abeta 1-42 significantly enhanced IL-1beta production in THP-1 monocytes. In contrast, a formyl chemotactic receptor antagonist inhibited Abeta 1-42-induced IL-1beta release from both human THP-1 monocytes and primary rat microglia. Further, primary rat microglia grown in culture expressed FPR as demonstrated by immunocytochemistry. Given the multiple pathophysiologic roles IL-1beta may play in AD, agents that block Abeta interactions with formyl chemotactic receptors on microglia might be important antiinflammatory therapeutic targets.

Alzheimer Disease↗

PubMedDate not supplied

Lack of HLA-molecules on human spermatozoa and in seminal plasma.

The expression of human leukocyte antigens (HLA) on ejaculated spermatozoa and on lymphocytes was compared by flow cytometry using monoclonal antibodies towards HLA class I (pan-HLA-A, -B, -C) and class II (DR) antigens. Soluble antigens of HLA class I (s HLA-A, -B, -C) in seminal plasma and in blood plasma were monitored with an ELISA technique. Lymphocytes showed specific fluorescence after incubation with the antibodies against HLA class I and class II (DR), whereas, on spermatozoa no positive immunofluorescence could be detected. No antibodies were bound to any significant extent either after modifications of sperm preparation (density gradient centrifugation, swim up-technique, addition of azide, foetal calf serum or benzamidine chloride) or after treatment of spermatozoa with detergents. Furthermore, different concentrations of soluble HLA-A, -B, -C in seminal plasma and in blood plasma were detected. The latter one showed soluble HLA about four-fold more concentrated than the seminal plasma (means +/- SD: 262.5 +/- 144.4 nmol l-1 vs. 62.5 +/- 27.1 nmol l-1). These results suggest, that the HLA-expression differs between human spermatozoa and somatic cells.

Flow Cytometry↗

PubMedDate not supplied

Disintegration of human spermatozoal membranes in seminal plasma decreases the binding capacity of integrins.

The in vitro binding capacity of spermatozoal integrins to matrix components after disintegration of sperm membranes was evaluated. The percentage of spermatozoa with functionally-relevant integrins was determined before and after devitalization of spermatozoa, which were resuspended in seminal plasma or in culture medium. The devitalization was performed by cryoshock or by incubation of spermatozoa with triton X-100 in a concentration ranging from 0.01 to 1.0%. The spermatozoal integrins were detected by the binding of anti-integrin antibodies and flow cytometry and the functional activity was monitored by the binding of the spermatozoa to the matrix components in a cell attachment assay. The seminal plasma decreased the binding of anti-integrin antibodies to the spermatozoal surface and the binding of spermatozoa to ligands and matrix components. respectively. In contrast, the expression of fibronectin and laminin on spermatozoa was increased. Not all spermatozoa, which expressed integrins on their surface bound to the ligands in the cell attachment assay. These results suggest that the detectable integrins only partially exert functional relevance. It can be concluded that the spermatozoa with fragile plasma membranes are more prone to functional inactivation of their integrins by the seminal plasma.

Cell Adhesion↗

PubMedDate not supplied

Localization of adhesion molecules on human spermatozoa by fluorescence microscopy.

The expression of adhesion molecules on human spermatozoa of healthy probands was analysed. The localization patterns of adhesion molecules (AM) on the spermatozoal surface were documented by fluorescence microscopy. Spermatozoa were incubated with antibodies against alpha 1 (CD49a), alpha 2 (CD49b), alpha 3 (CD49c), alpha 4 (CD49d), alpha 5 (CD49e), alpha 6 (CD49f) chains of beta 1 integrins, beta 1 (CD29), beta 2 (CD18), alpha V (CD51), beta 3 (CD61) and beta 4 integrin chains, the LFA-3 (Lymphocyte function antigen, CD58) from the immunoglobulin superfamily and the extra-cellular matrix proteins laminin, fibronectin and collagen IV. For collagen IV, alpha 1 and alpha 2 chains no expression could be noticed. Laminin was detected at the acrosomal membrane, fibronectin and beta 4 chain mainly at the equatorial membrane. The fibronectin receptors alpha 3, alpha 4 and alpha 5 chains of the beta 1 integrins were mainly located on acrosomal and equatorial membrane areas. Laminin receptor alpha 6 chain was located postacrosomal and less frequently acrosomal. beta 2 chain and vitronectin receptors alpha V and beta 3 chains had a mainly postacrosomal localization pattern. LFA-3 was found constantly on postacrosomal membrane areas. Double staining technique was used to prove the simultaneous occurrence of fibronectin and its integrin receptors alpha 3, alpha 4 and alpha 5 chains and of alpha V and beta 2 chains on spermatozoa. The localization patterns of integrins on double stained spermatozoa were similar to the patterns described for single stained spermatozoa. The localization of fibronectin appeared to be influenced by the presence of integrins: the typical equatorial fibronectin band disappeared in case of an equatorial localization of integrins.

Antigens, CD↗

PubMedDate not supplied

Adhesion molecules and matrix proteins on human spermatozoa.

Ejaculated spermatozoa and spermatogenic cells express alpha- and beta-chains of beta 1, 3 and 4 integrins as well as their ligands fibronectin and laminin in an extended intra- and interindividual variation and in different patterns of location. The mRNA transcripts of these molecules were detectable by nested polymerase chain reaction in the spermatozoa. The conclusion of a functional competence of these adhesion molecules (AM) was supported by their relation to the results of the zona-free hamster oocyte penetration (HOP) test, the in vitro fertilization of human oocytes and cell attachment assays. AM labelling was influenced by the disintegration of the sperm plasma membrane, especially in seminal plasma, by progesterone, human follicular fluid and microorganisms, but was barely modified by sperm cryopreservation. Despite substantial advances in the knowledge about sperm adhesion molecules, many questions remain to be answered.

Animals↗

PubMedDate not supplied

In vitro fertilization: increased VLA (very late antigen) integrins and fibronectin after acrosome reaction.

The pattern of detectable adhesion molecules (AM) on human ejaculated spermatozoa used for in vitro fertilization (IVF) of human oocytes was evaluated. The percentage of spermatozoa with the alpha chains 3, 4, 5, and 6 of the integrins (also called very late antigens, VLA alpha 3, 4, 5, 6), the alpha V and the beta 3 chains, as well as the matrix proteins laminin and fibronectin, were determined by flow cytometry using monoclonal antibodies against integrins and matrix proteins before and after the acrosome reaction. This reaction was induced by the low-temperature method according to P. Sanchez, E. Töpfer-Petersen, R. J. Aitken, and W. B. Schill (1991, Andrologia 23:197-203) and monitored by the binding of FITC-conjugated Pisum sativum agglutinin. Twenty-three of 34 investigated semen samples fertilized at least one oocyte (fertile group) and 11 samples did not induce a pronucleus formation or cleavage of an oocyte (infertile group). Significant differences in the classical semen parameters between both semen sample groups could be shown only for the percentage of progressively motile spermatozoa (p = .046, U test). The spermatozoa of the fertile group showed a significantly higher expression of VLA alpha 3 (p = .045) and fibronectin (p = .048). Additionally, after the loss of the acrosome a significantly higher expression of alpha 4, alpha 5, and alpha 6 chains of integrins (p < .05) was detected in the fertile semen group. In contrast, the inducibility of the acrosome reaction differed only to a nonsignificant extent (p = .094). These data suggest that spermatozoa showing a higher expression of the alpha chains of the integrins after the acrosome reaction have a better fertilization rate in vitro.

Acrosome↗

PubMedDate not supplied

Inhibition of tumor growth by plasminogen-related protein-B.

BACKGROUND: Various fragments of the fibrinolytic protein plasminogen can act as antiangiogenic factors and inhibit the growth of primary and metastatic tumors in mice. Plasminogen-related gene-B encodes a putative 9 kDa protein virtually identical to the plasminogen N-terminal activation peptide, a 77-amino acid motif that is liberated from the parent plasminogen molecule during conversion to the serine proteinase plasmin. Previous data have documented enhanced transcription of plasminogen-related gene-B in neoplastic tissues. MATERIALS AND METHODS: We have tested the effects of recombinant versions of plasminogen-related protein-B and the plasminogen N-terminal activation peptide on the growth of tumors in mice, employing murine tumor cell lines implanted subcutaneously. RESULTS: The recombinant plasminogen-related protein-B significantly inhibited the growth of primary tumors in mice, while recombinant plasminogen N-terminal activation peptide elicited only a slight inhibition of tumor growth. CONCLUSION: These data suggest that plasminogen-related protein-B may have utility as a novel cancer therapeutic.

Angiogenesis Inhibitors↗