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PubMed2005

Clinical, immunophenotypic, and genetic analysis of adult lymphomas with morphologic features of Burkitt lymphoma.

A prompt distinction of Burkitt lymphoma (BL) versus diffuse large B cell lymphomas (DLBCL) has important clinical implications; however, this distinction can be difficult. We analyzed 74 adult gray zone and 10 reference pediatric BL using immunohistochemistry (Ki-67, CD10, bcl2, bcl6) and fluorescence in situ hybridization (FISH) for MYC, BCL2, and BCL6 breakpoints. Two algorithms for classification were followed: algorithm A used a two-step review by four hemato-pathologists and algorithm B a set of only biologic markers (Ki-67 > or = 90%, CD10+, bcl6+, bcl2-, MYC breakpoint+, BCL2 and BCL6 breakpoint-). Both algorithms categorized all reference cases as BL. In the adult group, algorithm A resulted in 21 adult BL and 52 DLBCL and algorithm B in 23 BL and 51 "non-Burkitt" lymphomas (nBL); 9 cases (12%) contained two different translocations and were categorized as nBL in algorithm B. Fifteen cases (20%) fulfilled the BL criteria of both algorithms. Although not considered as BL according to both algorithms, many other lymphomas showed nonetheless a phenotypic and/or genetic shift to BL. BL according to algorithm B was more homogeneous with respect to clinical presentation (gender and localization) than BL defined by algorithm A. Our data suggest that only a few cases of these gray zone lymphomas represent true de novo BL. Immunohistochemistry for Ki-67, CD10, and bcl2 with analysis of MYC and preferably also BCL2 and BCL6 may be advised as a marker panel for this diagnostic dilemma.

Adolescent↗
PubMed1985

Comparison of the various forms of glutathione S-transferase with glucose-6-phosphate dehydrogenase and gamma-glutamyltranspeptidase as markers of preneoplastic and neoplastic lesions in rat kidney induced by N-ethyl-N-hydroxyethylnitrosamine.

Alterations in gamma-glutamyl transpeptidase (gamma-GT) and alkaline phosphatase (ALP) activities and binding of specific antibodies to glucose-6-phosphate dehydrogenase (G6PDH), gamma-GT and the A, B, C, D, and P forms of glutathione S-transferase (GST-A, -B, -C, -D, and -P, respectively) in preneoplastic and neoplastic lesions induced by N-ethyl-N-hydroxyethylnitrosamine (EHEN) in the rat kidney were investigated. Morphologically the lesions were of basophilic type and were classified either as altered tubules or adenomas, the latter being further subdivided into microadenomas and adenomas depending on size and the presence of compression. The fact that all lesions demonstrated only weak (or negative) gamma-GT, ALP and GST-B stainings, all of which are normally evident in proximal tubules, and also the occasional finding of cells bearing a periodic acid-Schiff-positive apical brush border within altered tubules strongly positive for GST-A, indicated their histogenesis from the proximal segment of the nephron. Thus a directed shift to a phenotype opposite to that observed in the cells of origin was demonstrated. Comparison of phenotype within the range of EHEN-induced lesions strongly suggested putative preneoplastic character for the altered tubules. Transition to adenomas appeared to be correlated with progressive loss of GST-A and moderate to slight increase of G6PDH.

Adenoma↗
PubMed1994

Alkaline phosphatase activity in cultured cells of human healthy and hyperplastic gingivae: a preliminary study.

The present study compared the alkaline phosphatase (ALP) expression in cultured cells derived from different types of gingival hyperplasia. Tissue biopsies were obtained from patients with clinically non-inflamed, enlarged gingivae of idiopathic gingivofibromatosis (IGF) and phenytoin-induced hyperplasia (PHG). Fibroblasts obtained from the gingivae or periodontal ligaments (PDL) of the same or different healthy subjects were used as controls. Fibroblasts were plated and the ALP activity and total amount of DNA per well were studied in each isolated cell type at 4, 7 and 9 days of culture. The results revealed that fibroblasts derived from IGF tissue synthesized a large amount of ALP by day 7 and day 9 of culture. The amount of ALP activity was cell density-dependent and the strongest expression of enzyme activity was noted after cell confluence. In contrast, only very low ALP activity was detected in cells derived from normal gingivae and PHG. Cytochemical examination of these cells revealed similar observations. The fact that fibroblasts derived from PHG tissue did not exhibit ALP in vitro suggests that there was a shift of cell phenotype in this tissue. The increased expression of ALP in IGF cells may imply a developmental trend toward osteoblastic phenotypes.

Alkaline Phosphatase↗
PubMed1995

The role of viral diversity in HIV pathogenesis.

From the time of seroconversion, patients who retain a high viral load are known to progress more rapidly to AIDS than individuals who have lower levels of HIV. Increased viral load is accompanied by decreasing CD4 cell numbers, and CD4 cell counts are therefore widely used as a diagnostic marker or predictor of progression in HIV patients. During the asymptomatic phase, however, CD4 counts often remain in the "normal" range, and this marker provides little information on potential disease progression. A shift in viral phenotype from non-syncytium-inducing (NSI) to syncytium-inducing (SI) heralds disease progression, and early in infection SI progression can be distinguished from nonprogression on the basis of CD4 changes. NSI progression, on the other hand, cannot be distinguished from nonprogression on the basis of CD4 counts but can be distinguished on the basis of RNA load. Molecular assessment of viral phenotype, such as quantitative measures of HIV RNA, as well as syncytium-inducing and nonsyncytium-inducing is therefore able to provide valuable information on the probable course of progression in individual patients, thus aiding in more effective disease management.

Acquired Immunodeficiency Syndrome↗
PubMed1996

Wild type neu transgene counteracts mutant homologue in malignant transformation of rat Schwann cells.

Mutational activation of the neu (erbB-2) receptor protein tyrosine kinase gene appears to be the triggering event in the process of oncogenesis induced by N-ethyl-N-nitrosourea (EtNU) in immature Schwann cells of the rat peripheral nervous system. Subsequent loss of the wild-type neu allele may represent a critical secondary step towards malignancy. Developmentally-regulated expression of a wild-type rat neu transgene (neu cDNA under the control of the rat Po promoter) in the Schwann cells of transgenic BDIX and Sprague-Dawley rats exposed to EtNU on postnatal day 1 results in a lower incidence of early atypical proliferates in the trigeminal nerve. Furthermore, re-introduction of the wild-type neu gene into homozygous neu mutant schwannoma cells counteracts the expression of the tumorigenic phenotype. The suppressive action of the wild-type gene over its mutationally activated oncogenic homologue underlines the critical function of the neu gene in the control of differentiation in the Schwann cell lineage, and provides evidence for the responsiveness of cellular phenotypes towards quantitative shifts in the dosage of wild-type vs mutant signal transducing molecules.

Alleles↗
PubMed2000

T-lymphocyte subpopulations do not express identical combinations of interleukin-2 receptor chains in the early phase of their activation and proliferation.

Owing to the contrasting observations in the field of interleukin(IL)-2 receptor research, the expression of IL-2 receptor chains was analysed on resting and anti-CD3 antibody (OKT3) activated CD4 and CD8 T cells by flow cytometry. Prior to the stimulation, 49% of CD4+ cells expressed IL-2Ralpha (CD25), whereas the expression of IL-2Rbeta (CD122) was very low (8%). The reverse was true for CD8 cells: 48% of them were positive for CD122, but only a fraction (10%) expressed CD25. Practically all lymphocytes expressed IL-2Rgamma (CD132). Interestingly, the unbalanced expression of IL-2Ralpha and -beta continued throughout the stimulation period of 2 days. In addition, the expression of CD45 isoforms in combination with the IL-2R chains and CD71 was followed during the activation of CD4+ T cells. Although CD45RA+/RO- CD4 cells were effectively activated, they retained their naive phenotype up to 2 days of stimulation. On the other hand, CD45RA+low/RO+low (Ddull) CD4+ cells shifted to the memory phenotype rapidly after being activated. However, by day 6 of stimulation the shift of both naive and Ddull cells to memory ones was obvious. The role of the IL-2 receptor in the activation of CD4 subpopulations is discussed.

Biomarkers↗
PubMed2000

Fine-tuning the EBV+ hu-PBL-SCID xenogeneic chimera model using in vivo superinfection.

Our purpose was to establish a reproducible xenogeneic chimera model to observe tumors similar to the well-known human posttransplant lymphoproliferative disease (LPD). First we followed the original protocol injecting Epstein-Barr virus positive (EBV+) human peripheral blood lymphocytes (PBL) intraperitoneally into immunodeficient (SCID) mice. Human cells showed T cell phenotype in majority one week after the transfer, whereas one month later a shift towards B cell phenotype was evident according to immunohistochemical and flow cytometric analysis. At this stage the intraperitoneal mass of cells suggested a biologically malignant behaviour infiltrating the liver and the spleen of the host animal. Immunohistochemistry indicated proliferating human lymphatic cells expressing EBV associated proteins and characteristic patterns of invasion within the affected organs. Eventually LPD was lethal to the host animals in 46-67 days. However, the microscopic appearance of experimental LPD was different from the human haemopoietic malignancies: the basic structures of lymphatic organs were preserved and the human T and B cells repopulated the normally T and B dependent areas in mice. The phenotypes of the proliferating cells were human and characteristic for the mature T- and B-lymphocytes. No dominant clone developed during in vitroculturing of the biologically invasive mass of cells removed from the tumor-bearing mice. The results of microscopical, immunological, and flow cytometrical analysis suggested a mature but uncontrolled proliferation of human lymphocytes in SCID mice. The original method for the induction of post-transplant LPD in SCID mice was modified in our further experiments to standardise the experimental technique increasing the efficiency of B cell proliferation and the reducing the number of unspecific factors. Subsequent in vivo EBV superinfection was carried out after the intraperitoneal transfer of a reduced quantity of human PBL from different donors. The same disease developed in our modified chimera model as by the use of original protocol except for some valuable differences. All hosts developed LPD regardless the significantly reduced amount of transplanted PBL and it was lethal in a shorter period of time (41-43 days) compared to the original model. The decreased quantity of transplanted human lymphatic cells was formerly insufficient using the original protocol. Therefore this modified and standardised protocol can lead to a more predictable and reproducible model allowing us to examine fine details of posttransplant lympho-proliferative disease.

Animals↗
PubMed2000

Interspecific Competition, Environmental Gradients, Gene Flow, and the Coevolution of Species' Borders.

Darwin viewed species range limits as chiefly determined by an interplay between the abiotic environment and interspecific interactions. Haldane argued that species' ranges could be set intraspecifically when gene flow from a species' populous center overwhelms local adaptation at the periphery. Recently, Kirkpatrick and Barton have modeled Haldane's process with a quantitative genetic model that combines density-dependent local population growth with dispersal and gene flow across a linear environmental gradient in optimum phenotype. To address Darwin's ideas, we have extended the Kirkpatrick and Barton model to include interspecific competition and the frequency-dependent selection that it generates, as well as stabilizing selection on a quantitative character. Our model includes local population growth, movements over space, natural selection, and gene flow. It simultaneously addresses the evolution of character displacement and species borders. It reproduces the Kirkpatrick and Barton single-species result that limited ranges can be produced with sufficiently steep environmental gradients and strong dispersal. Further, in the absence of environmental gradients or barriers to dispersal, interspecific competition will not limit species ranges at evolutionary equilibrium. However, interspecific competition can interact with environmental gradients and gene flow to generate limited ranges with much less extreme gradient and dispersal parameters than in the single-species case. Species display character displacement in sympatry, yet the reduction in competition that results from this displacement does not necessarily allow the two species to become sympatric everywhere. When species meet, competition reduces population densities in the region of overlap, which, in turn, intensifies the asymmetry in gene flow from center to margin. This reduces the ability of each species to adapt to local physical conditions at their range limits. If environmental gradients are monotonic but not linear, the transition zone between species at coevolutionary equilibrium occurs where the environmental gradient is steepest. If productivity gradients are also introduced into the model, then patterns similar to Rapoport's rule emerge. Interacting species respond to climate change, as it affects the optimal phenotype over space, by a combination of range shifts and local evolution in mean phenotype, while solitary species respond solely by range shifts. Finally, we compare empirical estimates for intrinsic growth rates and diffusion coefficients for several species to those needed by the single-species model to produce a stable limited range. These empirical values are generally insufficient to produce limited ranges in the model suggesting a role for interspecific interactions.

Journal Article↗
PubMed1993

Ia- macrophages and cytokine networks contribute to tumor-induced suppression of CD4+ autoreactive T cells.

Tumor growth changes the functions and phenotypes of macrophages (M phi) and T cells. Suppression of CD4+ T cell autoresponses during tumor growth was contributed primarily by M phi. Tumor-induced alterations in the abilities of these cells to mediate autorecognition were assessed through syngeneic mixed lymphocyte reaction (SMLR) assays. Tumor-bearing host (TBH) M phi were significantly more suppressive (60-90%) than normal host (NH) M phi, and this suppression was caused partly by reduced Ia expression. TBH Ia- M phi were significantly more suppressive (50-80%) than their NH counterparts. The suppression mechanism was controlled partly by prostaglandin E2 (PGE2), because treating cultures with indomethacin and titrated NH and TBH Ia- M phi led to increased T-cell responsiveness, although responsiveness never reached levels of assays containing unseparated M phi. Blocking studies using anti-interferon-gamma (anti-IFN-gamma) monoclonal antibodies (mAb), anti-interleukin 4 (anti-IL-4) mAb, and indomethacin suggested that IFN-gamma, IL-4, and PGE2 contributed to tumor-induced M phi-mediated suppression. Our results suggested that a quantitative shift in M phi phenotype and a qualitative shift in M phi function in addition to differences in cytokine-directed accessory activities are partly responsible for tumor-induced suppression CD4+ T cell autoresponses.

Animals↗
PubMed1982

Partial characterization of temperature-sensitive mutants of foot-and-mouth disease virus, O1 caseros strain.

The preliminary characterization of four temperature-sensitive (ts) mutants of the O1 Caseros strain of foot-and-mouth disease virus is described. Two mutants, ts 6 and ts 40, showed a very low RNA synthesis rate at nonpermissive temperature and were classified phenotypically as RNA(-). Shift-up experiments demonstrated an incapacity to organize the RNA replicative process at nonpermissive temperature. Another mutant (ts 139) behaved phenotypically as RNA(+) and its virions were more thermolabile than the wild type virus, so the defect in this mutant is likely to be in one of its structural proteins. Finally, mutant ts 5 was phenotypically RNA(+); its leak production at nonpermissive temperature was high, and the shift-up experiment showed a defect that was expressed either late in the replicative cycle or continuously.

Animals↗
PubMed1997

The second finger of Urbs1 is required for iron-mediated repression of sid1 in Ustilago maydis.

The urbs1 gene encodes a transcriptional regulator of siderophore biosynthesis in Ustilago maydis. Biological and DNA-binding activities of the two putative zinc-finger motifs of Urbs1 were studied by analyzing mutants containing altered finger domains. The mutated urbs1 alleles from three previously described N'-methyl-N'-nitro-N-nitrosoguanidine (NTG) mutants were mapped and cloned by a gap-repair procedure. Sequence analyses revealed single amino acid substitutions in two of the NTG mutants. Both mutations (G-507 to D in urbs1-1 and P-491 to L in urbs1-3), which are located in the Urbs1 C-terminal finger domain, reduced DNA-binding activity by 10-fold and were sufficient to confer a urbs1-minus phenotype. The third NTG urbs1 mutant (urbs1-2) also contained a mutation in one of the conserved amino acids (P-518 to S) in the C-terminal finger domain, but this mutation alone was not sufficient to confer a urbs1-minus phenotype. A second frame shift mutation was identified in urbs1-2 and is necessary for the urbs1-minus phenotype. In an analysis of the function of the N-terminal finger of Urbs1, the conserved amino acid Arg-350 was mutated to leucine. A Urbs1 protein with this mutation complemented a urbs1 null mutant strain. By contrast, a similar mutation in the C-terminal domain abolished the ability of Urbs1 to regulate siderophore biosynthesis and greatly reduced its ability to bind target DNA.

Amino Acid Sequence↗
PubMed2006

Ecological opportunity and phenotypic plasticity interact to promote character displacement and species coexistence.

We investigated the roles of resource availability and phenotypic plasticity in promoting ecological character displacement (i.e., trait evolution stemming from resource competition between species). Because ecological character displacement generates new populations that differ in resource use, this process should only occur when exploitable resources are available. We tested this hypothesis in two species of spadefoot toads (Spea bombifrons and S. multiplicata) whose tadpoles use phenotypic plasticity to develop into either an omnivore morph, which specializes on detritus, or a physically distinctive carnivore morph, which specializes on shrimp. Both species grow best on shrimp, but when reared together, S. bombifrons outcompetes S. multiplicata for shrimp and S. multiplicata outcompetes S. bombifrons for detritus. We found that when each species occurred alone in the field, they produced similar proportions of omnivores and carnivores. When the two species occurred together, however, they underwent ecological character displacement in larval development, with S. multiplicata producing mostly omnivores, and S. bombifrons producing mostly carnivores. We combined observations of natural populations with experiments to evaluate whether such character displacement was only possible when both shrimp and detritus were relatively abundant. Mixed-species ponds contained abundant detritus and shrimp, in contrast with nearby pure-species ponds, which were deficient in one resource. Experiments revealed that S. multiplicata competed poorly when detritus was rare and that S. bombifrons competed poorly when shrimp was rare. In nature, when one of these two resources was scarce, one species was missing, perhaps through competitive exclusion by the species that was the superior competitor for the remaining resource. Thus, ecological character displacement and, therefore, coexistence of close competitors, was only possible when diverse resources were available. Finally, even if exploitable resources are available, character displacement is not guaranteed to transpire if species cannot utilize such resources expeditiously. Phenotypic plasticity provides a general and important mechanism for facilitating resource partitioning. Thus, by facilitating shifts in resource use, phenotypic plasticity and ecological opportunity may often interact to promote divergence and coexistence of competitors.

Adaptation, Physiological↗
PubMedDate not supplied

Multiple phenotypic divergence of mammary adenocarcinoma cell clones. I. In vitro and in vivo properties.

The properties of cell clones derived from locally growing and spontaneous metastases of 13762NF mammary adenocarcinoma change during in vitro growth. This has been termed phenotypic drift and is reproducible in independent experiments using different cryoprotected cell stocks. To determine whether phenotypic drift in 13762NF cell clones is the result of an en bloc shift in the properties of all tumor cells, or independent phenotypic divergence of tumor cells to produce a mixed cell population, local tumor-derived clone MTF7 was subcloned at low and high culture passage numbers in vitro. Each subclone was analyzed in vitro for cell morphology, growth rate, saturation density, karyotype and ploidy, and in vivo for experimental metastatic behavior. Subclones derived from low passage clone MTF7 (T11; tissue culture passage number 11) were relatively homogeneous in their growth rates (doubling times of 16.8-17.4 h) and saturation densities (approximately 2 X 10(5) cells/cm2); yet, these same subclones were heterogeneous in their in vitro cell morphologies, experimental metastatic potentials (means range from 0 to greater than 100 tumor nodules per lung), size distributions of lung tumor nodules, marker chromosomes and modal chromosome numbers. High passage MTF7 (T35; tissue culture passage number 35) subclones had similar growth rates and saturation densities, except for subclone 2, which had a doubling time of approximately 26 h. Cell morphologies, experimental metastatic potentials (means range from 3 to greater than 600 tumor nodules per lung), size distribution of lung tumor nodules, marker chromosomes and modal chromosome numbers varied between MTF7 (T35) subclones. The results suggest that simultaneous, independent divergence of several phenotypes from a single cloned cell occurred to form a mixed cell population containing cells with independently segregated, unrelated phenotypes. Thus, the reproducibility of phenotypic drift in clonal cell populations was probably the result of tumor cell divergence and was not an en bloc shift in phenotypic properties of all cells.

Adenocarcinoma↗
PubMed1995

Stress-induced reduction in the rat mixed lymphocyte reaction is due to macrophages and not to changes in T cell phenotypes.

Exposure to aversive events or stressors modulates various aspects of immune function. We have previously reported that exposure to an acute stressor, inescapable tail shock (IS), resulted in a shift in T cell subpopulations in rat mesenteric lymph nodes but not in cervical lymph nodes (Fleshner et al. (1992) J. Neuroimmunol. 41, 131-142). The mesenteric CD4+/CD8+ ratio was increased immediately after exposure to IS and was due primarily to an increase in the percent of CD4+ cells. The present experiments were designed to determine the relationship between the IS-associated phenotypic shift and its significance in the function of CD4+ T cells. The function assessed was the in vitro proliferative response to alloantigens coded for by the Major Histocompatibility Complex (MHC). Using the mixed lymphocyte reaction (MLR), we report that exposure to IS resulted in a decrease in the MLR response of cells from both cervical and mesenteric lymph nodes. Depletion of macrophages (nylon wool adherent cells) eliminated the IS-induced reduction and co-culture of macrophages (irradiation-insensitive cells) from shocked rats produced the suppression. One interpretation of these data is that exposure to IS resulted in the activation of macrophages and the release of a suppressive factor which reduced the MLR response of peripheral lymph node lymphocytes.

Animals↗
PubMed1983

The interaction between daughterless and sex-lethal in triploids: a lethal sex-transforming maternal effect linking sex determination and dosage compensation in Drosophila melanogaster.

Regulation of Drosophila sex determination and X-chromosome dosage compensation in response to the X-chromosome/autosome (X/A) balance of the zygote is shown to require proper functioning of both the da+ gene in the mother and the Sxl+ gene in the zygote. Previous studies led to the hypothesis that zygotic Sxl+ alleles are differentially active in females (XXAA) vs males (XYAA) in response to the X/A balance, and that maternal da+ gene product acts as a positive regulator in this connection. Sxl+ activity was proposed to impose the female developmental sequence on cells which would follow the male sequence in its absence. Important predictions of this proposal are verified. This study focuses primarily on the phenotype of triploid intersexes (XXAAA, X/A = 0.67). They are shown here to survive effects of da and Sxl mutations that would be lethal to diploids. The ambiguous X/A signal of intersexes normally causes them to develop as phenotypic mosaics of male and female tissue. Loss of maternal da+ or zygotic Sxl+ gene function shifts their somatic sexual phenotype to the male alternative. A gain-of-function mutation at Sxl has the opposite effect, imposing female development regardless of the maternal genotype with respect to da. It also reduces their rate of X-linked gene expression. The effects of a duplication of Sxl+ resemble those of the constitutive Sxl allele, but are less extreme. The role of these genes in the process of X-chromosome dosage compensation is inferred indirectly from the strict dependence of the mutations' lethal effects on the X/A balance in haploids, diploids, and triploids, and more directly from the effects of the mutations on the phenotypes of the X-linked neomorphic mutations, Bar and Hairy-wing. The relationship of da+ and Sxl+ gene functions to those of other sex-specific lethal loci in D. melanogaster, and to sex determination mechanisms in other species, is discussed.

Animals↗
PubMed1998

Smoke-induced inhalation injury: effects of retinoic acid and antisense oligodeoxynucleotide on stability and differentiated state of the mucociliary epithelium.

Rabbit tracheal explants, exposed to burning pine wood smoke, were cultured in a chemically defined medium with and without retinoic acid (+/- RA). Exposures of 15-20 minute led to RA-independent degeneration of the mucociliary epithelial sheath. In 10 minute exposures tissue integrity was retained, but epithelial morphology changed from normal pseudostratified columnar to the flattened appearance typical of the squamous phenotype. Despite the dramatic shift in morphology, explants exhibited normal RA-dependent mucin gene expression characteristic of the mucociliary phenotype. Furthermore, electron micrographs showed continued presence of both secretory granules and cilia. RA(+) cultures also showed a normal pattern of adherent epithelial cells. In RA(-) cultures, however, there were prominent intercellular spaces indicating an RA dependence for maintaining adhesive contacts following smoke exposure. An 18-mer mucin antisense oligomer that suppressed mucin gene expression also unexpectedly blocked the smoke induced metaplasia in RA(+) cultures, but the sense oligomer had no effect.

Animals↗
PubMed1992

Evolution of the V3 envelope domain in proviral sequences and isolates of human immunodeficiency virus type 1 during transition of the viral biological phenotype.

The third variable domain (V3) of the envelope gene of human immunodeficiency virus type 1 contains a major neutralization epitope and determinants of syncytium-inducing (SI) capacity and replication rate (reviewed by J. P. Moore and P. L. Nara, AIDS Suppl. 2:S21-S33, 1991). Sequences were generated from DNA of samples taken 3 months apart over a period of 24 and 30 months from peripheral blood mononuclear cells (PBMC) of two individuals, both before and after cocultivation with uninfected donor PBMC. The isolated virus shifted from the non-syncytium-inducing (NSI) phenotype to the SI phenotype during the study period. This shift was associated with distinct changes in the V3 domain in both patients. The association of the phenotype shift with the V3 sequence changes was confirmed by construction of viruses with chimeric V3 loops. The shift from NSI- to SI-associated V3 variants was also seen in the uncultured PBMC of both patients, but not until 3 and 9 months after the detection of SI virus in culture. In the samples of uncultured PBMC DNA, several subgroups of sequences were found, indicating that the process of evolution may not be gradual and that several distinct populations can coexist. The paucity of intermediate sequences indicated that strong selection pressure was exerted on this part of the envelope. The early emergence of disease-associated SI variants in cultured material indicates that virus culture may have relevance for the in vivo situation.

Amino Acid Sequence↗
PubMed1993

"Trans-differentiation" from epidermal to mesenchymal/myogenic phenotype is associated with a drastic change in cell-cell and cell-matrix adhesion molecules.

Cells of the human keratinocyte line HaCaT were shifted to a mesenchymal/myogenic phenotype (DTHMZ cells) by MyoD1 transfection, 5-aza-2' deoxycytidine treatment, and selection for reduced adhesion on plastic. Since this correlated with loss of stratification (inability to form a multilayered tissue), we determined the status of cell-cell and cell-matrix adhesion molecules involved in epidermal morphogenesis. Expression of desmosomal proteins (plakoglobin, desmoglein, desmoplakin) and uvomorulin was no longer detectable at the mRNA and protein level in the DTHMZ cells while both HaCaT cells and malignant variants (transfected with c-Ha-ras oncogene) expressed uvomorulin in vitro and in transplants in vivo, the latter even in invasively growing tumor nodules. Furthermore, HaCaT cells stained positive for the integrin subunits beta 1, alpha 2, alpha 3, and alpha 5, typical for cultured keratinocytes. In contrast, the putative fibronectin receptor alpha 5 beta 1, common also in fibroblasts, was the only integrin showing strong staining in DTHMZ cells. The integrin subunits alpha v and a6, clearly expressed at the mRNA level, weakly stained HaCaT cultures and led to a dotlike fluorescence in DTHMZ cells, possibly representing focal adhesion plaques. The respective integrin status correlated well with the growth behavior on different matrices. While HaCaT cells readily attached and proliferated on collagen (type I), fibronectin-coated, and laminin-coated collagen gels, DTHMZ cells formed monolayers only on fibronectin-coated collagen. This was, however, not sufficient to allow stratification in vivo. Altogether, the status of adhesion molecules in DTHMZ cells more likely reflects that seen in mesenchymal cells as compared to the pattern of keratinocytes displayed by HaCaT cells. Moreover, since the DTHMZ cells were clearly HaCaT descendants, the results support our hypothesis of a "trans-differentiation" process from an epidermal (HaCaT) to a mesenchymal/myogenic phenotype (DTHMZ).

Cadherins↗