Minor histocompatibility antigens HA-1-, -2-, and -4-, and HY-specific cytotoxic T-cell clones inhibit human hematopoietic progenitor cell growth by a mechanism that is dependent on direct cell-cell contact

Abstract HLA-identical bone marrow transplantation (BMT) may be complicated by graft-versus-host disease or graft rejection. Both complications are thought to be initiated by recognition of minor histocompatibility (mH) antigens by HLA-restricted mH-antigen-specific T lymphocytes. Using HLA- A2-restricted mH antigens HA-1-, -2-, and -4-, and HY-specific cytotoxic T lymphocyte (CTL) clones, we studied the recognition by these CTL clones of interleukin-2 (IL-2)-stimulated T cells (IL-2 blasts), BM mononuclear cells (BMMNCs), and hematopoietic progenitor cells (HPCs). We showed that, when IL-2 blasts from the BM donors who were investigated were recognized by the HA-1-, -2-, and -4-, and HY- specific CTL clones, their BMMNCs and HPCs were recognized as well by these CTL clones, resulting in antigen-specific growth inhibition of erythrocyte burst-forming units (BFU-E), colony-forming units- granulocyte (CFU-G), and CFU-macrophage (CFU-M). the HA-2-specific CTL clone, however, inhibited BFU-E and CFU-G growth from four donors to a lesser extent than from two other donors. We further investigated whether inhibitory cytokines released into the culture medium by the antigen-specific stimulated CTLs or by stimulated BMMNCs were responsible for suppression of HPC growth or whether this effect was caused by direct cell-cell contact between CTLs and HPCs. HPC growth inhibition was only observed after preincubation of BMMNCs and CTLs together for 4 hours before plating the cells in semisolid HPC culture medium. When no cell-cell contact was permitted before plating, neither antigen-stimulated CTL nor antigen-nonstimulated CTLs provoked HPC growth inhibition. Culturing BMMNCs in the presence of supernatants harvested after incubation of BMMNCs and CTL clones together for 4 or 72 hours did also not result in HPC growth inhibition. Both suppression of HPC growth and lysis of IL-2 blasts and BMMNCs in the 51Cr-release assay appeared to be dependent on direct cell-cell contact between target cells and CTLs and were not caused by the release of inhibitory cytokines into the culture medium by antigen-specific stimulated CTLs or by stimulated BMMNCs. Our results show that mH-antigen-specific CTLs can inhibit HPC growth by a direct cytolytic effect and may therefore be responsible for BM graft rejection after HLA-identical BMT.

Download Full-text

Minor histocompatibility antigens HA-1-, -2-, and -4-, and HY-specific cytotoxic T-cell clones inhibit human hematopoietic progenitor cell growth by a mechanism that is dependent on direct cell-cell contact

Blood ◽

10.1182/blood.v82.12.3778.bloodjournal82123778 ◽

1993 ◽

Vol 82 (12) ◽

pp. 3778-3785 ◽

Cited By ~ 1

Author(s):

WA Marijt ◽

WF Veenhof ◽

E Goulmy ◽

R Willemze ◽

JJ van Rood ◽

...

Keyword(s):

Growth Inhibition ◽

Graft Rejection ◽

Culture Medium ◽

Hematopoietic Progenitor Cell ◽

Cytotoxic T Cell ◽

Cell Contact ◽

Target Cells ◽

Hematopoietic Progenitor ◽

Direct Cell ◽

Cell Cell

HLA-identical bone marrow transplantation (BMT) may be complicated by graft-versus-host disease or graft rejection. Both complications are thought to be initiated by recognition of minor histocompatibility (mH) antigens by HLA-restricted mH-antigen-specific T lymphocytes. Using HLA- A2-restricted mH antigens HA-1-, -2-, and -4-, and HY-specific cytotoxic T lymphocyte (CTL) clones, we studied the recognition by these CTL clones of interleukin-2 (IL-2)-stimulated T cells (IL-2 blasts), BM mononuclear cells (BMMNCs), and hematopoietic progenitor cells (HPCs). We showed that, when IL-2 blasts from the BM donors who were investigated were recognized by the HA-1-, -2-, and -4-, and HY- specific CTL clones, their BMMNCs and HPCs were recognized as well by these CTL clones, resulting in antigen-specific growth inhibition of erythrocyte burst-forming units (BFU-E), colony-forming units- granulocyte (CFU-G), and CFU-macrophage (CFU-M). the HA-2-specific CTL clone, however, inhibited BFU-E and CFU-G growth from four donors to a lesser extent than from two other donors. We further investigated whether inhibitory cytokines released into the culture medium by the antigen-specific stimulated CTLs or by stimulated BMMNCs were responsible for suppression of HPC growth or whether this effect was caused by direct cell-cell contact between CTLs and HPCs. HPC growth inhibition was only observed after preincubation of BMMNCs and CTLs together for 4 hours before plating the cells in semisolid HPC culture medium. When no cell-cell contact was permitted before plating, neither antigen-stimulated CTL nor antigen-nonstimulated CTLs provoked HPC growth inhibition. Culturing BMMNCs in the presence of supernatants harvested after incubation of BMMNCs and CTL clones together for 4 or 72 hours did also not result in HPC growth inhibition. Both suppression of HPC growth and lysis of IL-2 blasts and BMMNCs in the 51Cr-release assay appeared to be dependent on direct cell-cell contact between target cells and CTLs and were not caused by the release of inhibitory cytokines into the culture medium by antigen-specific stimulated CTLs or by stimulated BMMNCs. Our results show that mH-antigen-specific CTLs can inhibit HPC growth by a direct cytolytic effect and may therefore be responsible for BM graft rejection after HLA-identical BMT.

Download Full-text

Faculty Opinions recommendation of Direct cell-cell contact with the vascular niche maintains quiescent neural stem cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.719710075.793506153 ◽

2015 ◽

Author(s):

Mirna Perez-Moreno

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Cell Contact ◽

Vascular Niche ◽

Direct Cell ◽

Cell Cell

Download Full-text

Abstract 15392: Inflammation and Ischemic Heart Failure: Monocyte-Mediated Cardiac Fibroblast Activation and Matrix Remodeling Through a Direct Cell-Cell Contact Mechanism

Circulation ◽

10.1161/circ.130.suppl_2.15392 ◽

2014 ◽

Vol 130 (suppl_2) ◽

Author(s):

Holly E Mewhort ◽

Brodie D Lipon ◽

Daniyil A Svystonyuk ◽

David G Guzzardi ◽

Paul W Fedak

Keyword(s):

Peripheral Blood ◽

Cardiac Fibroblast ◽

Cell Contact ◽

Β1 Integrin ◽

Blood Monocytes ◽

Peripheral Blood Monocytes ◽

Fibroblast Activation ◽

Direct Cell ◽

Tgf Β1 ◽

Cell Cell

BACKGROUND: Following myocardial infarction (MI), activated cardiac myofibroblasts facilitate extracellular matrix (ECM) remodeling to prevent mechanical complications. However, prolonged myofibroblast activity leads to dysregulation of the ECM, maladaptive remodeling, fibrosis and heart failure (HF). Chronic inflammation is believed to drive persistent myofibroblast activity, however, the mechanisms are unclear. In this study, we explored the effects of peripheral blood monocytes on human cardiac fibroblast activation in a 3D ECM microenvironment. METHODS/RESULTS: Human cardiac fibroblasts isolated from surgical human heart biopsies were seeded into 3D collagen matrices. Peripheral blood monocytes isolated from healthy human donors were co-cultured with fibroblasts. Monocytes increased fibroblast activation measured by collagen ECM contraction (17.9±11.1% increase; p<0.01) and resulted in local ECM remodeling observed by confocal microscopy. Under co-culture conditions that prevent cell-cell contact but allow interaction via paracrine factors, monocytes had minimal effects on fibroblast activation (6.4±7.0 vs.17.9±11.1% increase, respectively; p<0.01). Multiplex analysis of the co-culture media revealed an increase in the paracrine factors Transforming Growth Factor-beta 1 (TGF-β1) and Matrix Metalloproteinase 9 when monocytes and fibroblasts were cultured under cell-cell contact conditions (162.2±11.7pg/mL and 17.5±0.5ng/mL, respectively, vs. 21.8±5.7pg/mL and 4.9 ±0.4ng/mL; p<0.001). TGF-β1 blockade abolished monocyte induced cardiac fibroblast activation, as did β1-integrin. These data suggest direct cell-cell interaction between monocytes and cardiac fibroblasts through β1-integrin results in TGF-β1 release facilitating fibroblast activation and matrix remodeling. CONCLUSION: For the first time, we demonstrate that peripheral blood monocytes stimulate human cardiac fibroblast activation through a mechanism involving TGF-β1 release as a consequence of direct cell-cell interaction through β1-integrin. These data implicate inflammation as a driver of cardiac fibrosis post-MI, highlighting potential novel therapeutic targets for the treatment of ischemic HF.

Download Full-text

RNAi-Mediated Knock-Down of Arylamine N-acetyltransferase-1 Expression Induces E-cadherin Up-Regulation and Cell-Cell Contact Growth Inhibition

PLoS ONE ◽

10.1371/journal.pone.0017031 ◽

2011 ◽

Vol 6 (2) ◽

pp. e17031 ◽

Cited By ~ 40

Author(s):

Jacky M. Tiang ◽

Neville J. Butcher ◽

Carleen Cullinane ◽

Patrick O. Humbert ◽

Rodney F. Minchin

Keyword(s):

Growth Inhibition ◽

Cell Contact ◽

Knock Down ◽

E Cadherin ◽

Cell Cell

Download Full-text

CD26/Dipeptidylpeptidase IV Regulates Potency of Selected Hematopoietic Growth Factors through Truncation, and Recovery In Vivo After Cytotoxic Stress.

Blood ◽

10.1182/blood.v114.22.3602.3602 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3602-3602

Author(s):

Hal E. Broxmeyer ◽

Jonathan Hoggatt ◽

Scott Cooper ◽

Giao Hangoc ◽

Louis M. Pelus ◽

...

Keyword(s):

Stem Cell ◽

Colony Formation ◽

Hematopoietic Progenitor Cell ◽

Target Cells ◽

Hematopoietic Progenitor ◽

Dipeptidylpeptidase Iv ◽

Gm Csf ◽

Hematopoietic Recovery ◽

Stimulation Of

Abstract Abstract 3602 Poster Board III-539 CD26 is a dipeptidylpeptidase IV (DPPIV) that cleaves dipeptides from the N-terminus after a proline or alanine. An amino acid sequence search identified putative CD26/DPPIV truncation sites in a number of colony stimulating factors (CSFs), including human (hu) and mouse (mu) GM-CSF and G-CSF, hu IL-3, and hu and mu EPO. These truncation sites were not apparent in mu IL-3, hu and mu M-CSF, or in hu and mu stem cell factor (SCF) or Flt3-ligand (FL). We hypothesized that CD26/DPPIV served as a regulator of the potency of the selected CSFs that contain this putative truncation site, and that hematopoietic recovery after stress would be enhanced and accelerated in CD26 −/− mice. We first used Diprotin A (Ile-Pro-Ile), a known CD26/DPPIV inhibitor for mu and hu cells. Mu cytokines were assessed for activity on mu BM, and hu cytokines on hu cord blood (CB), all in dose-response fashion. Hu EPO was tested on mu and hu cells. One hour pre-treatment of mu BM cells with Diprotin A, or use of CD26 −/− mu BM cells, resulted in a two-fold or greater enhancement of CFU-GM-, CFU-G-, and BFU-E- colony formation of cells respectively stimulated by mu GM-CSF, mu G-CSF, and hu EPO. The CSF activities of mu M-CSF for CFU-M, and mu IL-3 for CFU-GM were not enhanced by inhibition/deletion of CD26/DPPIV. Also, pretreatment of hu CB cells with Diprotin A, enhanced colony formation of CFU-GM stimulated by hu GM-CSF or hu IL-3, and of BFU-E stimulated by hu EPO, but did not influence stimulation of CFU-G or CFU-M by hu M-CSF. Stimulation of cells with two CSFs results in additive to greater than additive hematopoietic progenitor cell (HPC) colony formation compared to that of each CSF alone. When both CSFs had putative CD26/DPPIV truncation sites, colony formation by HPC was even further increased by pretreatment of target cells with Diprotin A. Pretreatment of cells with Diprotin A did not enhance colony formation of mu BM or hu CB cells each respectively stimulated with SCF or FL alone, nor did it enhance the synergistic effects noted when SCF or FL was used in combination with CSFs. To assess truncation and activity directly, CSFs were pretreated with purified soluble DPPIV. Truncation was detected by Mass Spectrometry for CSFs with the putative truncation site, but not for those without this site. Truncated CSFs manifested greatly reduced activity against target cells, effects that were more apparent when the cells were pretreated with Diprotin A. Moreover, mixture of a truncated CSF with a non-truncated form of that CSF reduced colony formation to the level of the truncated CSF, suggesting that the truncated CSF interfered with or blocked activity of the full-length CSF. To evaluate effects of CD26/DPPIV on hematopoietic recovery, CD26 −/− and +/+ mice were treated with sublethal dosages of irradiation, or either cycle-specific or non-cycle specific drugs. In all cases, enhanced and accelerated recovery of hematopoietic progenitor cells was noted in the CD26 −/− mice, compared to the control CD26 +/+ mice. These results demonstrate that CD26/DPPIV regulates the activity of selected CSFs, and inhibition/deletion of CD26/DPPIV allows for enhanced in vitro potency of selected CSFs, and in vivo recovery of hematopoiesis in mice stressed with irradiation and cytotoxic drugs. These results may have practical relevance for understanding, and manipulating hematopoietic recovery after cytotoxic treatment or hematopoietic stem cell transplantation. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

INFLAMMATION INDUCES POST-ISCHEMIC HEART FAILURE: SYSTEMIC MONOCYTES STIMULATE HUMAN CARDIAC MYOFIBROBLAST ACTIVATION BY DIRECT CELL-CELL CONTACT

Canadian Journal of Cardiology ◽

10.1016/j.cjca.2014.07.231 ◽

2014 ◽

Vol 30 (10) ◽

pp. S155

Author(s):

H.E. Mewhort ◽

B.D. Lipon ◽

D.A. Svystonyuk ◽

D.G. Guzzardi ◽

P.W. Fedak

Keyword(s):

Heart Failure ◽

Ischemic Heart ◽

Cell Contact ◽

Ischemic Heart Failure ◽

Direct Cell ◽

Cell Cell

Download Full-text

Mutual Enhancement of Differentiation of Osteoblasts and Osteocytes Occurs Through Direct Cell-Cell Contact

Journal of Cellular Biochemistry ◽

10.1002/jcb.24880 ◽

2014 ◽

pp. n/a-n/a ◽

Cited By ~ 4

Author(s):

Koji Fujita ◽

Qian Xing ◽

Sundeep Khosla ◽

David G. Monroe

Keyword(s):

Cell Contact ◽

Direct Cell ◽

Cell Cell

Download Full-text

Endocytic Motif on a Biotin-Tagged HIV-1 Env Modulates the Co-Transfer of Env and Gag during Cell-to-Cell Transmission

Viruses ◽

10.3390/v13091729 ◽

2021 ◽

Vol 13 (9) ◽

pp. 1729

Author(s):

María Inés Barría ◽

Raymond A. Alvarez ◽

Kenneth Law ◽

Deanna L. Wolfson ◽

Thomas Huser ◽

...

Keyword(s):

Fluorescent Imaging ◽

Productive Infection ◽

Live Cells ◽

Cell Contact ◽

Target Cells ◽

Structural Protein ◽

Cell Infection ◽

Virological Synapses ◽

Hiv 1 ◽

Cell Cell

During HIV-1 transmission through T cell virological synapses, the recruitment of the envelope (Env) glycoprotein to the site of cell–cell contact is important for adhesion and for packaging onto nascent virus particles which assemble at the site. Live imaging studies in CD4 T cells have captured the rapid recruitment of the viral structural protein Gag to VSs. We explored the role of endocytic trafficking of Env initiated by a membrane proximal tyrosine motif during HIV transfer into target cells and examined the factors that allow Gag and Env to be transferred together across the synapse. To facilitate tracking of Env in live cells, we adapted an Env tagging method and introduced a biotin acceptor peptide (BAP) into the V4 loop of Env gp120, enabling sensitive fluorescent tracking of V4-biotinylated Env. The BAP-tagged and biotinylated HIVs were replication-competent in cell-free and cell-to-cell infection assays. Live cell fluorescent imaging experiments showed rapid internalized cell surface Env on infected cells. Cell–cell transfer experiments conducted with the Env endocytosis mutant (Y712A) showed increased transfer of Env. Paradoxically, this increase in Env transfer was associated with significantly reduced Gag transfer into target cells, when compared to viral transfer associated with WT Env. This Y712A Env mutant also exhibited an altered Gag/biotin Env fluorescence ratio during transfer that correlated with decreased productive cell-to-cell infection. These results may suggest that the internalization of Env into recycling pools plays an important role in the coordinated transfer of Gag and Env across the VS, which optimizes productive infection in target cells.

Download Full-text