Cytochemical studies of the vascular endothelium

Cytochemical methods have been used to examine the vascular endothelium. With hemeproteins and immunocytochemistry, investigators have demonstrated the pathways that blood-borne molecules can take to gain access to the extravascular space (Ghitescu et al. 1986; Milici et al. 1987; Schneeberger and Karnovsky 1971; Simionescu et al. 1975). These same cytochemical methods have also provided evidence that morphologically similar endothelia may have different permeability properties (Hart and Pino 1985b, 1986; Pino 1985; Pino and Essner 1980, 1981). Differences in the location and chemical composition of cell surface moieties have been ascertained with enzyme digestion methods, lectins, and cationic ferritin (De Bruyn and Michelson 1978; Pino 1984c, 1986a, b; Simionescu et al. 1981a). The author hopes that he has provided the reader with representative examples of how investigators have used these cytochemical methods for their studies. As new methods are developed and applications are found for existing techniques such as ultracryomicrotomy (Milici et al. 1987) and colloidal gold markers (Pino 1987b), cytochemistry will remain a fundamental tool for the study of the structure and function of the vascular endothelium.     

Efficient algorithms for searching for exact repetition of nucleotide sequences

Summary There are several algorithms designed for searches for homologous sequences (Fitch 1966; Needleman and Wunsch 1970; Chva'tal and Sankoff 1975; Griggs 1977; Sannkoff 1972; Smith and Waterman 1981; Smith et al. 1981, Wagner and Fisher 1974; Waterman et al. 1976). This paper presents some very simple and useful high speed, text editing algorithms that search for exact nucleotide sequence repetition and genome duplication. The last algorithm suggested here is specifically adapted for the 4-letter alphabet of nucleotide sequences. Owing to the rapid accumulation of nucleotide sequences and the frequent need to search for sequence repetition or where a given set of nucleotides occurs in long sequences, efficient algorithms of this type are a necessity.     

Primary and secondary structure of U2 snRNA

With the improved rapid sequencing techniques, the earlier sequence of U2 RNA of Novikoff hepatoma (Shibata et al, J. Biol. Chem. 250, 3909-3920, 1975) was reanalyzed and modified. The improved sequence of U2 RNA is 188 (or 189) nucleotides long and is in register with a characterized U2 RNA pseudogene (Denison et al, PNAS 78, 810-814, 1981) except for an 11 nucleotide sequence (nucleotides 147-157) which is absent from the pseudogene. From these results, a secondary structure of U2 RNA is proposed which is supported by the preferred cleavage sites with T1-RNase, RNase A and S1 nuclease. Isolated U2 RNA was cleaved by T1-RNase preferentially at positions 64 and 164, whereas U2 RNA in U2-snRNP was cleaved only at position 64, indicating that position 164 is protected in U2-snRNP. As with U1 RNA (Epstein et al, PNAS 78, 1562-1566, 1981) the 5'-end of isolated U2 RNA was not preferentially cleaved by T1-RNase.     

Resistance of Adenoviral DNA Replication to Aphidicolin Is Dependent on the 72-Kilodalton DNA-Binding Protein

Aphidicolin is a highly specific inhibitor of DNA polymerase α and has been most useful for assessing the role of this enzyme in various replication processes (J. A. Huberman, Cell 23:647-648, 1981). Both nuclear DNA replication and simian virus 40 DNA replication are highly sensitive to this drug (Krokan et al., Biochemistry 18:4431-4443, 1979), whereas mitochondrial DNA synthesis is completely insensitive (Zimmerman et al., J. Biol. Chem. 255:11847-11852, 1980). Adenovirus DNA replication is sensitive to aphidicolin, but only at much higher concentrations. These patterns of sensitivity are seen both in vivo and in vitro (Krokan et al., Biochemistry 18:4431-4443, 1979). A temperature-sensitive mutant of adenovirus type 5 known as H5ts125 is able to complete but not initiate new rounds of replication at nonpermissive temperatures (P. C. van der Vliet and J. S. Sussenbach, Virology 67:415-426, 1975). When cells infected with H5ts125 were shifted from permissive (33°C) to nonpermissive (41°C) conditions, the residual DNA synthesis (elongation) showed a striking increase in sensitivity to aphidicolin. The temperature-sensitive mutation of H5ts125 is in the gene for the 72-kilodalton single-stranded DNA-binding protein. This demonstrated that the increased resistance to aphidicolin shown by adenovirus DNA replication was dependent on that protein. It also supports an elongation role for both DNA polymerase α and the 72-kilodalton single-stranded DNA-binding protein in adenovirus DNA replication. Further support for an elongation role of DNA polymerase α came from experiments with permissive temperature conditions and inhibiting levels of aphidicolin in which it was shown that newly initiated strands failed to elongate to completion.     

Sulfite binding to a flavodehydrogenase, cytochrome b2 from baker's yeast.

Baker's yeast L-lactate dehydrogenase (flavocytochrome b2) is a typical flavodehydrogenase, in that it accepts two electrons from the substrate but has a monoelectronic acceptor. Yet it forms a red semiquinone [Capeillère Blandin et al. Eur. J. Biochem. 54, 549--566 (1975)] and it is shown in this paper that it forms a reversible covalent complex with sulfite (Kd = 1.4 muM). This complex can be observed by difference spectroscopy and provides a convenient tool for visualizing the flavin chromophore, usually hidden behind the intense heme absorbance. A number of anions (D-lactate, oxalate and pyruvate) are inhibitors of the enzymatic reaction and induce spectral perturbations of the flavin spectrum. It is concluded that probably two positive charges exist at the active site: one which stabilizes the red semiquinone and one which attracts organic anions and sulfite. It is also concluded that the correlation between reactivity with sulfite and reactivity with oxygen among flavo-proteins may not be as general as previously proposed [Massey et al. J. Biol. Chem. 244, 3999--4006 (1969)].     

Waiting to inhale: HIF-1 modulates aerobic respiration

The hypoxia-inducible factor HIF-1 is known to promote anaerobic respiration during low oxygen conditions (hypoxia). In this issue, Fukuda et al. (2007) expand the range of HIF-1's functions by showing that it modulates aerobic respiration as well.     

Antipain microinjection prevents progesterone to inhibit adenyl cyclase in Xenopus oocytes

Microinjection of antipain, an inhibitor of thiol and Ca2+-dependent proteases, in immature Xenopus oocytes inhibited meiotic maturation induced by progesterone, but not by transfer of cytoplasm taken from maturing oocytes. Oocytes could be released from antipain inhibition by increasing progesterone concentration. alpha-32P-ATP was microinjected to study adenylcyclase in ovo. As already reported, neosynthesis of cAMP was decreased following progesterone application. This decrease was not observed, or it was considerably reduced, in oocytes previously injected with antipain. In amphibian, full-grown ovarian oocytes are arrested at first meiotic prophase, and have a large nucleus known as the germinal vesicle. Progesterone induces the production of a cytoplasmic maturation-promoting factor (MPF), which itself triggers germinal vesicle breakdown (GVBD), and subsequent events of meiotic maturation (Masui and Markert, 1971; Gerhart et al., 1984). A considerable body of evidences support the view that release from prophase block is due to inactivation of a cAMP-dependent protein kinase (reviewed by Maller, 1983). On the other hand, progesterone has been shown to induce a transient decrease in cAMP level (Speaker and Butcher, 1977; Schorderet-Slatkine et al., 1982; Cicirelli et al., 1985), and this initial drop of cAMP, along with a number of studies indicating a decrease in adenylate cyclase activity (Mulner et al., 1979; Baltus et al., 1981; Sadler and Maller, 1981; Finidori-Lepicard et al., 1981; Jordana et al., 1981), provided key support to the theory that an early drop in cAMP led to the dephosphorylation of a hypothetical protein which initiates maturation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cloning and nucleotide sequence of the structural gene encoding for human tryptophanyl-tRNA synthetase.

A structural gene encoding bovine (b) tryptophanyl-tRNA synthetase (WRS) has recently been cloned and sequenced [Garret et al., Biochemistry 30 (1991) 7809-7817]. Using part of this sequence as a hybridisation probe we have cloned and sequenced a structural gene encoding human polypeptide highly homologous with two mammalian proteins, bWRS [Garret et al., Biochemistry 30 (1991) 7809-7817; EMBL accession No. X52113] and rabbit peptide chain release factor [Lee et al., Proc. Natl. Acad. Sci. USA 87 (1990) 3508-3512]. Identification of the sequence encoding a human WRS is based on (i) the presence of 'HIGH' and 'KMSKS' structural motifs typical for class-I aminoacyl-tRNA synthetases [Eriani et al., Nature 347 (1990) 203-206]; (ii) coincidence of the number of SH groups per subunit estimated experimentally [Muench et al., Science 187 (1975) 1089-1091] and deduced from the cDNA sequence (six in both cases); (iii) close resemblance of two WRS polypeptides sequenced earlier [Muench et al., Science 187 (1975) 1089-1091] and the predicted structure in two different regions.     

Deletion mapping of kil and kor functions in the trfA and trfB regions of broad host range plasmid RK2

Figurski et al. (1982) have reported that certain loci on the broad host range plasmid RK2 (kil functions) can be cloned only in the presence of other trans-acting segments of the plasmid genome (kor functions). They have suggested that the presence of these functions may in part account for the structure of mini RK2 replicons which were constructed in order to define the regions of the plasmid which encode replication/maintenance functions (Thomas et al. 1980). We have therefore investigated the relationship between these two sets of kil and kor loci and the loci implicated in the replication/maintenance of RK2. We find that, whilst the three kil loci reported by Figurski et al. (1982) are absent from these derivatives, a fourth such locus (kilD) is closely linked to trfA, a gene essential for RK2 replication. The kilD locus was probably responsible for the inclusion in mini replicons of a segment of RK2 DNA which carries both korD and korA in addition to trfB, a gene defined by a temperature-sensitive maintenance defect, but which can be deleted leaving a functional RK2 replicon (Thomas 1981 b). The kilB locus is situated on the opposite side of kilD from trfA, all three loci lying within a 3.6 kb segment of RK2 DNA. The korA, korD and trfB functions all map within a 900 bp segment of DNA, while korB requires sequence information at least 1.5 kb from this segment.     

Quantitative microspectrophotometrical determination of protein thiols and disulfides with 2,2'-dihydroxy-6,6'-dinaphthyldisulfide (DDD). The variety of DDD-staining methods demonstrated on Ehrlich ascites tumor cells

2,2'-dihydroxy-6,6'-dinaphthyldisulfide (DDD) reacts with both protein thiol groups and with protein disulfides (N?hammer 1977). By varying the pH of the DDD-reaction, as well as the reaction times, the complex reaction became specific with respect to the histochemical demonstration of protein-SH groups. Furthermore, the application of the histochemical DDD-reaction following quantitative blockade of the protein-SH groups enabled the demonstration of distinctive DDD-reactive disulfides. The specificity and the extent of the different histochemical DDD-staining methods were investigated by comparing macroscopically determined values of the protein-SH-contents, and the contents of the different kinds of disulfides in Ehrlich-ascites-tumor cells (EATC) (Modig 1968; Hofer 1975), with microspectrometrical values determined with the MCN-method of N?hammer et al. (1981), and with microspectrometrical values measured on EATC after staining with the modified DDD-methods. Also, the method for the histochemical demonstration of protein-SH with DDD after the reduction of the disulfides with thioglycolate was investigated and conditions were found by which the protein-SH content could be determined quantitatively with DDD and Fast blue B after the reduction of the disulfides. With the aid of the MCN-method (N?hammer et al. 1981), the intracellular disulfide interchange reaction was investigated, leading to pH-dependent changes of the SH-SS-ratio of fixed cells during their incubation in aqueous media. In addition the possibility of protein loss during the long incubation times of the fixed cells in the DDD-solutions was investigated. For the quantitative microscpecrometrical determination of the protein content of EATC the so-called tetrazonium-coupling method, optimized by N?hammer (1978) and calibrated by N?hammer et al. (1981), was used.     

Soybean glycinin A1aB1b subunit has a molecular chaperone-like function to assist folding of the other subunit having low folding ability

Soybean (Glycine max L.) glycinin is composed of five subunits which are classified into two groups (group I: A1aB1b, A1bB2, and A2B1a; group II: A3B4 and A5A4B3). All the common soybean cultivars contain both group I and II subunits (Maruyama, N. et al., Phytochemistry, 64, 701-708 (2003)). The biosynthesis of group I starts earlier compared with that of the A3B4 subunit during seed development (Meinke, D.W. et al., Planta, 153, 130-139 (1981)). We have revealed that group I A1aB1b was mostly expressed as a soluble protein, but that A3B4 was expressed mainly as an insoluble protein in Escherichia coli under the same expression conditions; namely, A1aB1b had higher folding ability than A3B4. We therefore assumed that A1aB1b assists folding of group II subunits like a molecular chaperone does. In order to ascertain this, A1aB1b and A3B4 were co-expressed in E. coli. All of the expressed proteins of A3B4 were recovered in a soluble fraction. To confirm this result, we also co-expressed A1aB1b with modified A3B4 versions having extremely low folding ability. All expressed modified A3B4 versions were soluble. These results clearly suggest that A1aB1b has a molecular chaperone-like function in their folding.     

Atomic coordinates and molecular conformation of yeast phenylalanyl tRNA. An independent investigation.

The atomic coordinates of yeast tRNA(Phe) in the monoclinic crystal form have been determined by an independent analysis from a model built into a 3 A MIR map. The overall molecular structure is found to be in agreement with those reported for the same crystal form by Ladner et al. (1975) and for the orthorhombic form by Quigley et al. (1975) and Kim et al. (1975). However, significant differences between any two of the four models are found in certain local regions of the molecule. The structure is analyzed in terms of the nucleotide stereochemistry and internucleotide phosphodiesters. A striking observation is that the majority of the nucleotide moieties occur in the conformation preferred by the constituent mononucleotides themselves. The internucleotide P-O bonds afford the primary source of flexibility for the folding of the polynucleotide backbone while the sugar pucker and C(4')-C(5') torsions provide the secondary source of flexibility.     

Plant transformation by microinjection techniques

Several techniques have been developed for introducing cloned genes into plant cells. Vectorless delivery systems such as PEG-mediated direct DNA uptake (e.g. Pasz-kowski et al. 1984), electroporation (e.g. Shillito et al. 1985), and fusion of protoplasts with liposomes (Deshayes et al. 1985) are routinely used in many experiments (see several chapters of this issue). A wide range of plant species, dicotyledonous as well as monocotyledonous, has been transformed by these vectorless DNA transfer systems. However, the availability of an efficient protoplast regeneration system is a prerequisite for the application of these techniques. For cells with intact cell walls and tissue explants the biological delivery system of virulent Agrobacterium species has been routinely used (for review see Fraley et al. 1986). However, the host range of Agrobacterium restricts the plant species, which can be transformed using this vector system. In addition, all these methods depend on selection systems for recovery of transformants. Therefore a selection system has to be established first for plant species to be transformed. The microinjection technique is a direct physical approach, and therefore host-range independent, for introducing substances under microscopical control into defined cells without damaging them. These two facts differentiate this technique from other physical approaches, such as biolistic transformation and macroinjection (see chapters in this issue). In these other techniques, damaging of cells and random manipulation of cells without optical control cannot be avoided so far. In recent years microinjection technology found its application in plant sciences, whereas this technique has earlier been well established for transformation of animal tissue culture cells (Capecchi 1980) and the production of transgenic animals (Brin-ster et al. 1981, Rusconi and Schaffner 1981). Furthermore, different parameters affecting the DNA transfer via microinjection, such as the nature of microinjected DNA, and cell cycle stage, etc, have been investigated extensively in animal cells (Folger et al. 1982, Wong and Capecchi 1985), while analogous experiments on plant cells are still lacking.     

Histochemistry and functional significance of putative neocortical transmitter substances: a review

1. Intrinsic neuronal chains of the neocortex communicate most probably with amino acid transmitters. These involve both excitatory (glutamate, aspartate--Nadler et al. 1976) both inhibitory (GABA--Ribak 1978) amino acids, and ensure fast, ionotropic postsynaptic actions (Eccles, McGeer 1979). 2. Some interneurons of the neocortex seemingly operate with the peptide transmitter VIP (Lorén et al. 1979). Presumably, this is a metabotropic, slowly acting substance (Dodd, Kelly and Said 1979). 3. The existence of intrinsic cholinergic neurons in the neocortex is a matter of question (Krnjevic and Silver 1965). It is worth to mention that in the periphery, cholinergic terminals also contain and release VIP (H?kfelt et al. 1980). It is not known, whether this transmitter dualism can be found in neocortex, too. An ascending cholinergic system projecting from the basal forebrain to the neocortex exists and exerts profound influence on cortical function (Shute and Lewis 1967). 4. Diffusely terminating, ascending monoamine axons innervate the neocortex and modulate interneuronal transmission (Thiery et al. 1977; Morrison et al. 1981, Lidov et al. 1981). 5. The neuropeptide SP excites cortical neurons (Phillis and Limacher 1974), and its presence in thin axons can be demonstrated immunohistochemically (H?kfelt et al. 1976). 6. Neocortical efferents to the thalamus and striatum seemingly use glutamate or aspartate (Fonnum et al. 1981). The transmitters of other corticofugal projections are not known. 7. The transmitters of specific thalamic afferents and those of callosal and association projections are unknown, too. 8. The main task of future histochemistry is to explore the synaptology of neocortical neurons and afferent systems with identified or evidenced transmitters, viz. to explore the neurochemical subsystems of cortical organization. The tool for it could be the immunohistochemistry, and future development depends mainly on the synthesis and purification of suitable antigens. The knowledge on the synaptology of identified neurochemical units of the cortex would be the basis of the understanding at least partly of the pharmacological effects exerted by the putative neocortical transmitters.     

A theory of follicle selection: II. Computer simulation of estradiol administration in the primate

A theory of follicle selection (Lacker, 1981) is tested in the primate by simulating the effects of estradiol administration at different times, strengths, and durations during the follicular phase of the menstrual cycle (Clark et al., 1981; Zeleznik, 1981; Dierschke et al., 1985). The theory can account for the observed atretogenic effects of circulating estradiol on follicle development including full, partial, and delayed atresia of the dominant follicle (Dierschke et al., 1985) and can explain why similar estradiol doses achieve different qualitative effects when given at different times during the cycle. The theory predicts that recovery from early atresia may be possible, and it can also account for the loss of control in the number of maturing follicles that has been observed when estradiol antibodies are given in the midfollicular phase (Zeleznik et al., 1985). These results support the hypothesis that the selection mechanism in the primate is a consequence of feedback involving an essentially equipotent follicle population interacting through circulating estradiol and pituitary gonadotropins. A quantitative test of the theory awaits experimental identification of the maturation surfaces that are predicted by it. An experimental design for this purpose is proposed.     

Phosphorylation sites on phosphoprotein NS of vesicular stomatitis virus.

The phosphoprotein NS of vesicular stomatitis virus which accumulates within the infected cell cytoplasm is phosphorylated at multiple serine and threonine residues (G. M. Clinton and A. S. Huang, Virology 108:510-514, 1981; Hsu et al., J. Virol. 43:104-112, 1982). Using incomplete chemical cleavage at tryptophan residues, we mapped the major phosphorylation sites to the amino-terminal half of the protein. Analysis of phosphate-labeled tryptic peptides suggests that essentially all of the label is within the large trypsin-resistant fragment predicted from the sequence of Gallione et al. (J. Virol. 39:52-529, 1981). A similar result has been obtained for NS protein isolated from the virus particle by C.-H. Hsu and D. W. Kingsbury (J. Biol. Chem., in press). Analysis of phosphodipeptides utilizing the procedures of C. E. Jones and M. O. J. Olson (Int. J. Pept. Protein Res. 16:135-142, 1980) enabled us to detect as many as six distinct phosphate-containing dipeptides. From these studies, together with the known sequence data, we conclude that the major phosphate residues on cytoplasmic NS protein are located in the amino third of the NS molecule and most probably between residues 35 and 106, inclusive. The studies also provide formal chemical proof that NS protein has a structure consistent with a monomer of the sequence of Gallione et al. as modified by J. K. Rose (personal communication). The low electrophoretic mobility of this protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis is not therefore due to dimerization.     

Cross-circulation studies on the influence of hypoxia and hypoxaemia on neuro-epithelial bodies in young rabbits

Summary The reactions of the previously described neuro-epithelial bodies (NEB) (Lauweryns et al., 1969, 1970, 1972a, b, 1973a, b, c, 1974, 1975) in young rabbits to: (1) hypoxia with normoxaemia in the arteria pulmonalis on the one hand, and (2) hypoxaemia in the arteria pulmonalis with normoxic aeration on the other hand, has been investigated by means of cross-circulation experiments and light microscopical, electron microscopical and morphometrical techniques.Hypoxically aerated young rabbits, which received normoxaemic blood in their arteria pulmonalis from a donor rabbit by means of an arterio-arterial cross-circulation with mutual exchange transfusion, revealed an increased exocytosis of the dense-core vesicles of their NEB. Normoxically aerated young rabbits which received hypoxaemic blood in an identical manner, did not exhibit an increased exocytosis.It is concluded that the NEB apparently react directly to the hypoxia of the inhaled air and not to the hypoxaemia of the pulmonary blood. By the release of serotonin and a polypeptide substance, they may produce a local vasoconstriction in hypoxically aerated lung areas, enabling an intrapulmonary regulation of the V/Q ratio. This is regarded as additional proof that the NEB — while being modulated by the CNS — probably are intrapulmonary chemoreceptors with local secretory activities, reacting to the composition of the inhaled air.     

Morphological identification of on- and off-centre brisk transient (Y) cells in the cat retina

Brisk transient (Y) cells were recorded extracellularly in the cat retina. The position and shape of their receptive field centres were plotted on a tangent screen, together with retinal landmarks, such as blood vessels adjacent to the recording area. After recording the retina was processed as a whole mount and stained with a reduced-silver method (see appendix). This technique stains the entire alpha cell population including the dendritic trees. Alpha cells are the morphological correlate of the brisk transient cells (Boycott & W?ssle 1974; Cleland et al. 1975). Maps of the screen plot and the histological preparation could be accurately superimposed by means of the retinal landmarks and each recorded brisk transient unit could unequivocally be attributed to a particular alpha cell. Alpha cell dendritic trees are unistratified in either of two laminae within the inner plexiform layer: (1) close to the inner nuclear layer border, 'outer alpha cells', or (2) about 10 micrometers further towards the ganglion cell layer, 'inner alpha cells'. This stratification difference can be observed in whole mounts for large populations of cells (W?ssle et al. 1981). Of the recorded brisk transient cells, all on-centre units were inner alphas and all off-centre units outer alphas.     

Studying the Neural Basis of Adaptive Locomotor Behavior in Insects

Studying the neural basis of walking behavior, one often faces the problem that it is hard to separate the neuronally produced stepping output from those leg movements that result from passive forces and interactions with other legs through the common contact with the substrate. If we want to understand, which part of a given movement is produced by nervous system motor output, kinematic analysis of stepping movements, therefore, needs to be complemented with electrophysiological recordings of motor activity. The recording of neuronal or muscular activity in a behaving animal is often limited by the electrophysiological equipment which can constrain the animal in its ability to move with as many degrees of freedom as possible. This can either be avoided by using implantable electrodes and then having the animal move on a long tether (i.e. Clarac et al., 1987; Duch & Pflüger, 1995; Böhm et al., 1997; Gruhn & Rathmayer, 2002) or by transmitting the data using telemetric devices (Kutsch et al, 1993; Fischer et al., 1996; Tsuchida et al. 2004; Hama et al., 2007; Wang et al., 2008). Both of these elegant methods, which are successfully used in larger arthropods, often prove difficult to apply in smaller walking insects which either easily get entangled in the long tether or are hindered by the weight of the telemetric device and its batteries. In addition, in all these cases, it is still impossible to distinguish between the purely neuronal basis of locomotion and the effects exerted by mechanical coupling between the walking legs through the substrate. One solution for this problem is to conduct the experiments in a tethered animal that is free to walk in place and that is locally suspended, for example over a slippery surface, which effectively removes most ground contact mechanics. This has been used to study escape responses (Camhi and Nolen, 1981; Camhi and Levy, 1988), turning (Tryba and Ritzman, 2000a,b; Gruhn et al., 2009a), backward walking (Graham and Epstein, 1985) or changes in velocity (Gruhn et al., 2009b) and it allows the experimenter easily to combine intra- and extracellular physiology with kinematic analyses (Gruhn et al., 2006).We use a slippery surface setup to investigate the timing of leg muscles in the behaving stick insect with respect to touch-down and lift-off under different behavioral paradigms such as straight forward and curved walking in intact and reduced preparations.     

Role of ICAM-3 in Intercellular Adhesion and Activation of T Lymphocytes

The immune system requires a fine regulation of intercellular communication for its normal function. There are several regulated molecular pathways involved in leukocyte cell interactions (Springer, 1990; Hynes, 1992). Among them, the interaction of the leukocyte integrin LFA-1 (CDlla/CD18) with its ligands provides multiple accessory adhesion signals of capital importance during different functions of the immune response, such as antigen presentation (Harding and Unanue, 1991), T-B lymphocyte interaction (Moy and Brian, 1992), cellular cytotoxicity (Makgoba et al., 1988; Altmann et al., 1989; Davignon et al, 1981; Akella and Hall, 1992), allogenic and autologous mixed lymphocyte reactions (Bagnasco et al., 1990) and recirculation and homing of lymphocytes through tissue endothelium (Hamann et al., 1988; Pals et al, 1988).