Flow cytometric characterization of human umbilical cord blood lymphocytes: Immunophenotypic features

Giovanni D'Arena, Pellegrino Musto, Nicola Cascavilla, Girolamo Di Giorgio, Saverio Fusilli, Federica Zendoli, Mario Carotenuto

Research output: Contribution to journalArticle

119 Citations (Scopus)

Abstract

Background and Objective. One of the most important potential advantages in the use of human umbilical cord blood (HUCB) for hematopoietic reconstitution after myeloablative therapy seems to be the lower occurrence of acute graft-versus-host-disease (GvHD) in recipients after allogeneic transplantation. Since mature T cells play an important role in GvHD pathogenesis, we tried to verify whether a different immunophenotypic pattern exists between HUCB and peripheral blood (PB) T cells. Design and Methods. An immunophenotypic study on 40 HUCB and 40 PB samples from healthy adult volunteers was performed, by means of flow cytometry using a large panel of monoclonal antibodies in double labeling. Results. The absolute lymphocyte count was greater in HUCB (5233±1808 μL) than in adult PB (1941±378 μL). Significant differences in percentage were found between cord and adult T- cells, respectively (CD3+: 59.9±12 vs 74.9±4.6%), CD3-CD16+ and/or CD56+ natural killer (NK) cells (23.8±10.1 vs 10.8±5.3%) and CD3+CD16+ and/or CD56+ cytotoxic T lymphocyte subset (0.3±0.3 vs 10.7±4.1%). There was no difference in CD4/CD8 ratio (1.7±0.5 vs 1.6±0.2%) between the two groups. In absolute terms, HUCB contained an higher number of all lymphocyte subsets, with the exception of CD3+ CD16+ and/or CD56+ T lymphocyte subpopulation, CD3+ CD25+ and CD3+HLA-DR+ activated T-cells. CD38, a marker of activation and immaturity, was present on virtually all cord T cells and on approximately half of the adult T cells. The large majority of HUCB T cells co-expressed CD45RA naive antigen (CD4+CD45RA+: 87.6±5.2%, CD8+ CD45RA+: 93.5±7.8%; CD4+CD45RO+: 12.3±5.2%; CD8+CD45RO+: 6.4±7.8%) whereas in adult PB T cells an higher number of CD45RO+ memory cells was detected (CD4+CD45RA+: 44.8±9.6%; CD8+CD45RA+: 71.5%±8.1%; CD4+CD45RO+: 55.2±9.6%; CD8+ CD45RO+: 28.5±8.1%). Finally, less than 14% of lymphocytes were shown to belong to B lineage In both sources, while, in absolute terms, they were more represented in HUCB with respect to adult PB. Interpretation and Conclusions. In the present study we found significant differences between HUCB and adult PB lymphocytes in their immunophenotypic profile. In particular HUCB showed T lymphocytes that appeared to be phenotypically immature. Indeed, as a likely consequence of poor antigenic experience during pregnancy, the majority of HUCB cells were naive, expressing the RA isoform of the CD45 molecule. These findings could justify the previously reported reduced cord blood lymphocyte alloreactivity when allogoneic transplantation is performed and require further functional studies in order to confirm the impairment of HUCB immune system response to alloantigens.

Original languageEnglish
Pages (from-to)197-203
Number of pages7
JournalHaematologica
Volume83
Issue number3
Publication statusPublished - Mar 1998

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Fetal Blood
Lymphocytes
T-Lymphocytes
Blood Cells
Lymphocyte Subsets
Graft vs Host Disease
CD45 Antigens
CD4-CD8 Ratio
Isoantigens
Homologous Transplantation
Lymphocyte Count
T-Lymphocyte Subsets
Cytotoxic T-Lymphocytes
HLA-DR Antigens
Natural Killer Cells
Immune System
Healthy Volunteers
Flow Cytometry
Protein Isoforms
Cell Count

Keywords

  • Flow cytometry
  • Graft-versus-host disease
  • Human umbilical cord blood
  • Lymphocyte subpopulations
  • Transplantation

ASJC Scopus subject areas

  • Hematology

Cite this

D'Arena, G., Musto, P., Cascavilla, N., Di Giorgio, G., Fusilli, S., Zendoli, F., & Carotenuto, M. (1998). Flow cytometric characterization of human umbilical cord blood lymphocytes: Immunophenotypic features. Haematologica, 83(3), 197-203.

Flow cytometric characterization of human umbilical cord blood lymphocytes : Immunophenotypic features. / D'Arena, Giovanni; Musto, Pellegrino; Cascavilla, Nicola; Di Giorgio, Girolamo; Fusilli, Saverio; Zendoli, Federica; Carotenuto, Mario.

In: Haematologica, Vol. 83, No. 3, 03.1998, p. 197-203.

Research output: Contribution to journalArticle

D'Arena, G, Musto, P, Cascavilla, N, Di Giorgio, G, Fusilli, S, Zendoli, F & Carotenuto, M 1998, 'Flow cytometric characterization of human umbilical cord blood lymphocytes: Immunophenotypic features', Haematologica, vol. 83, no. 3, pp. 197-203.
D'Arena, Giovanni ; Musto, Pellegrino ; Cascavilla, Nicola ; Di Giorgio, Girolamo ; Fusilli, Saverio ; Zendoli, Federica ; Carotenuto, Mario. / Flow cytometric characterization of human umbilical cord blood lymphocytes : Immunophenotypic features. In: Haematologica. 1998 ; Vol. 83, No. 3. pp. 197-203.
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T1 - Flow cytometric characterization of human umbilical cord blood lymphocytes

T2 - Immunophenotypic features

AU - D'Arena, Giovanni

AU - Musto, Pellegrino

AU - Cascavilla, Nicola

AU - Di Giorgio, Girolamo

AU - Fusilli, Saverio

AU - Zendoli, Federica

AU - Carotenuto, Mario

PY - 1998/3

Y1 - 1998/3

N2 - Background and Objective. One of the most important potential advantages in the use of human umbilical cord blood (HUCB) for hematopoietic reconstitution after myeloablative therapy seems to be the lower occurrence of acute graft-versus-host-disease (GvHD) in recipients after allogeneic transplantation. Since mature T cells play an important role in GvHD pathogenesis, we tried to verify whether a different immunophenotypic pattern exists between HUCB and peripheral blood (PB) T cells. Design and Methods. An immunophenotypic study on 40 HUCB and 40 PB samples from healthy adult volunteers was performed, by means of flow cytometry using a large panel of monoclonal antibodies in double labeling. Results. The absolute lymphocyte count was greater in HUCB (5233±1808 μL) than in adult PB (1941±378 μL). Significant differences in percentage were found between cord and adult T- cells, respectively (CD3+: 59.9±12 vs 74.9±4.6%), CD3-CD16+ and/or CD56+ natural killer (NK) cells (23.8±10.1 vs 10.8±5.3%) and CD3+CD16+ and/or CD56+ cytotoxic T lymphocyte subset (0.3±0.3 vs 10.7±4.1%). There was no difference in CD4/CD8 ratio (1.7±0.5 vs 1.6±0.2%) between the two groups. In absolute terms, HUCB contained an higher number of all lymphocyte subsets, with the exception of CD3+ CD16+ and/or CD56+ T lymphocyte subpopulation, CD3+ CD25+ and CD3+HLA-DR+ activated T-cells. CD38, a marker of activation and immaturity, was present on virtually all cord T cells and on approximately half of the adult T cells. The large majority of HUCB T cells co-expressed CD45RA naive antigen (CD4+CD45RA+: 87.6±5.2%, CD8+ CD45RA+: 93.5±7.8%; CD4+CD45RO+: 12.3±5.2%; CD8+CD45RO+: 6.4±7.8%) whereas in adult PB T cells an higher number of CD45RO+ memory cells was detected (CD4+CD45RA+: 44.8±9.6%; CD8+CD45RA+: 71.5%±8.1%; CD4+CD45RO+: 55.2±9.6%; CD8+ CD45RO+: 28.5±8.1%). Finally, less than 14% of lymphocytes were shown to belong to B lineage In both sources, while, in absolute terms, they were more represented in HUCB with respect to adult PB. Interpretation and Conclusions. In the present study we found significant differences between HUCB and adult PB lymphocytes in their immunophenotypic profile. In particular HUCB showed T lymphocytes that appeared to be phenotypically immature. Indeed, as a likely consequence of poor antigenic experience during pregnancy, the majority of HUCB cells were naive, expressing the RA isoform of the CD45 molecule. These findings could justify the previously reported reduced cord blood lymphocyte alloreactivity when allogoneic transplantation is performed and require further functional studies in order to confirm the impairment of HUCB immune system response to alloantigens.

AB - Background and Objective. One of the most important potential advantages in the use of human umbilical cord blood (HUCB) for hematopoietic reconstitution after myeloablative therapy seems to be the lower occurrence of acute graft-versus-host-disease (GvHD) in recipients after allogeneic transplantation. Since mature T cells play an important role in GvHD pathogenesis, we tried to verify whether a different immunophenotypic pattern exists between HUCB and peripheral blood (PB) T cells. Design and Methods. An immunophenotypic study on 40 HUCB and 40 PB samples from healthy adult volunteers was performed, by means of flow cytometry using a large panel of monoclonal antibodies in double labeling. Results. The absolute lymphocyte count was greater in HUCB (5233±1808 μL) than in adult PB (1941±378 μL). Significant differences in percentage were found between cord and adult T- cells, respectively (CD3+: 59.9±12 vs 74.9±4.6%), CD3-CD16+ and/or CD56+ natural killer (NK) cells (23.8±10.1 vs 10.8±5.3%) and CD3+CD16+ and/or CD56+ cytotoxic T lymphocyte subset (0.3±0.3 vs 10.7±4.1%). There was no difference in CD4/CD8 ratio (1.7±0.5 vs 1.6±0.2%) between the two groups. In absolute terms, HUCB contained an higher number of all lymphocyte subsets, with the exception of CD3+ CD16+ and/or CD56+ T lymphocyte subpopulation, CD3+ CD25+ and CD3+HLA-DR+ activated T-cells. CD38, a marker of activation and immaturity, was present on virtually all cord T cells and on approximately half of the adult T cells. The large majority of HUCB T cells co-expressed CD45RA naive antigen (CD4+CD45RA+: 87.6±5.2%, CD8+ CD45RA+: 93.5±7.8%; CD4+CD45RO+: 12.3±5.2%; CD8+CD45RO+: 6.4±7.8%) whereas in adult PB T cells an higher number of CD45RO+ memory cells was detected (CD4+CD45RA+: 44.8±9.6%; CD8+CD45RA+: 71.5%±8.1%; CD4+CD45RO+: 55.2±9.6%; CD8+ CD45RO+: 28.5±8.1%). Finally, less than 14% of lymphocytes were shown to belong to B lineage In both sources, while, in absolute terms, they were more represented in HUCB with respect to adult PB. Interpretation and Conclusions. In the present study we found significant differences between HUCB and adult PB lymphocytes in their immunophenotypic profile. In particular HUCB showed T lymphocytes that appeared to be phenotypically immature. Indeed, as a likely consequence of poor antigenic experience during pregnancy, the majority of HUCB cells were naive, expressing the RA isoform of the CD45 molecule. These findings could justify the previously reported reduced cord blood lymphocyte alloreactivity when allogoneic transplantation is performed and require further functional studies in order to confirm the impairment of HUCB immune system response to alloantigens.

KW - Flow cytometry

KW - Graft-versus-host disease

KW - Human umbilical cord blood

KW - Lymphocyte subpopulations

KW - Transplantation

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