Blood Res.  2018 Mar;53(1):61-70. 10.5045/br.2018.53.1.61.

Expression of adhesion molecules on CD34+ cells from steady-state bone marrow before and after mobilization and their association with the yield of CD34+ cells

Affiliations
  • 1Oncologia Clínica e Experimental, Universidade Federal de São Paulo – UNIFESP, Sao Paulo, Brazil. kcecyn@gmail.com

Abstract

BACKGROUND
Cell adhesion molecules (CAMs) expressed on hematopoietic progenitor cells (HPCs), endothelial cells, and stromal cells play a pivotal role in the mobilization of CD34+ cells. Herein, we conducted a non-randomized peripheral blood stem cell (PBSC) mobilization study aimed to compare the potential differences in the expressions of several CAMs and chemokines on CD34+ cells obtained from bone marrow aspirate before and after HPC mobilization from patients with hematologic malignancies and healthy donors.
METHODS
Three-color cytofluorometric analysis was used to compare the expressions of CAMs and chemokines in the bone marrow before and after mobilization.
RESULTS
For all studied groups, CAM expression among those with good and poor yields of CD34+ cells was significantly correlated with VCAM-1 (P=0.007), CD44 (P=0.027), and VLA-4 (P=0.014) expressions. VCAM-1 (P=0.001), FLT-3 (P=0.001), CD44 (P=0.011), VLA-4 (P=0.001), and LFA-1 (P=0.001) expressions were higher before HPC mobilization than after HPC mobilization. By contrast, the expression of CXCR4 significantly varied before and after mobilization only among those with successful PBSC mobilization (P=0.002).
CONCLUSION
We attempted to identify particular aspects of CAMs involved in CD34+ cell mobilization, which is a highly complex mechanism that involves adhesion molecules and matrix metalloproteases. The mechanism by which CD34+ cell mobilization is activated through proteolytic enzymes is not fully understood. We believe that CXCR4, VLA-4, CD44, and VCAM-1 are the most important molecules implicated in HPC mobilization, particularly because they show a correlation with the yield of CD34+ cells collected via large volume leukapheresis.

Keyword

Adhesion molecules; Hematopoietic progenitor cells; Mobilization; Stem cell donor; Multiple myeloma; Non-Hodgkin lymphoma

MeSH Terms

Bone Marrow*
Cell Adhesion Molecules
Chemokines
Endothelial Cells
Hematologic Neoplasms
Hematopoietic Stem Cells
Humans
Integrin alpha4beta1
Leukapheresis
Lymphocyte Function-Associated Antigen-1
Lymphoma, Non-Hodgkin
Metalloproteases
Multiple Myeloma
Peptide Hydrolases
Stem Cells
Stromal Cells
Tissue Donors
Vascular Cell Adhesion Molecule-1
Cell Adhesion Molecules
Chemokines
Integrin alpha4beta1
Lymphocyte Function-Associated Antigen-1
Metalloproteases
Peptide Hydrolases
Vascular Cell Adhesion Molecule-1

Figure

  • Fig. 1 Representative flow cytometric scattergrams from bone marrow aspirate specimens after HPC mobilization showing a 3-color cytofluorometric analysis of the expression of adhesion molecule antigens on CD34+ cells population. (A) CD34+ cells are painted in the lympho-mononuclear region. (B) CD34PE+ cells are included in R1 in the FSC/SSC dot plot. (C) CD34PE+ cells population in R2. (D) CD34FITC+ cells are included in R1 in the FSC/SSC dot plot. (E) CD34FITC+ cells population in R3. (F) CD34PE+ cells are included in R6 in the CD45PerCP/SSC dot plot. (G) CD34FITC+ cells are included in R6 in the CD45PerCP/SSC dot plot.

  • Fig. 2 Representative flow cytometric histograms from bone marrow aspirate specimens after HPC mobilization showing a 3-color cytofluorometric analysis of the expression of adhesion molecule antigen on CD34+ cell population. The results were estimated as the mean fluorescence intensity (MFI). The histograms (A) and (B) are IgG1 FITC and PE controls. (C) CD106, (D) CD135, (E) CD184, (F) CD62L, (G) CD49d, (H) CD11a, and (I) CD44 antigen on CD34+ cell population. The gating protocols have been described before.

  • Fig. 3 The mean expression of CD106, CD-44, and CD49d on the CD34+ cells assessed in BMA before mobilization and its association with the yield of CD34+ cells collected via LVL in both donors and patients.

  • Fig. 4 Distribution of CD106, CD135, CD11a, CD44, CD49d, and CD184 assessed in mononuclear cells of the BMA, before and after mobilization according to the yield of CD34+ cells obtained via LGV.

  • Fig. 5 Results obtained through logistic regression analysis. The chance of poor mobilization increases by approximately 6% with each increase of one unit of CD49d and by approximately 4% with each increase of one unit of CD44 in the pre-mobilization phase.


Reference

1. Arslan O, Moog R. Mobilization of peripheral blood stem cells. Transfus Apher Sci. 2007; 37:179–185. PMID: 17980665.
Article
2. Ikeda K, Kozuka T, Harada M. Factors for PBPC collection efficiency and collection predictors. Transfus Apher Sci. 2004; 31:245–259. PMID: 15556472.
Article
3. Haverkos BM, McBride A, O'Donnell L, et al. An effective mobilization strategy for lymphoma patients after failed upfront mobilization with plerixafor. Bone Marrow Transplant. 2014; 49:1052–1055. PMID: 24797182.
Article
4. Angelopoulou MK, Tsirkinidis P, Boutsikas G, Vassilakopoulos TP, Tsirigotis P. New insights in the mobilization of hematopoietic stem cells in lymphoma and multiple myeloma patients. Biomed Res Int. 2014; 2014:835138. PMID: 25197663.
Article
5. Yuan S, Wang S. How do we mobilize and collect autologous peripheral blood stem cells? Transfusion. 2017; 57:13–23. PMID: 27731496.
Article
6. Olivieri A, Marchetti M, Lemoli R, et al. Proposed definition of ‘poor mobilizer’ in lymphoma and multiple myeloma: an analytic hierarchy process by ad hoc working group Gruppo ItalianoTrapianto di Midollo Osseo. Bone Marrow Transplant. 2012; 47:342–351. PMID: 21625224.
Article
7. Fruehauf S, Seggewiss R. It's moving day: factors affecting peripheral blood stem cell mobilization and strategies for improvement. Br J Haematol. 2003; 122:360–375. PMID: 12877663.
8. Kröger N, Zeller W, Hassan HT, Dierlamm J, Zander AR. Difference between expression of adhesion molecules on CD34+ cells from bone marrow and G-CSF-stimulated peripheral blood. Stem Cells. 1998; 16:49–53. PMID: 9474747.
9. Chan JY, Watt SM. Adhesion receptors on haematopoietic progenitor cells. Br J Haematol. 2001; 112:541–557. PMID: 11260052.
Article
10. Ford CD, Greenwood J, Anderson J, Snow G, Petersen FB. CD34+ cell adhesion molecule profiles differ between patients mobilized with granulocyte-colony-stimulating factor alone and chemotherapy followed by granulocyte-colony-stimulating factor. Transfusion. 2006; 46:193–198. PMID: 16441594.
Article
11. Oostendorp RA, Reisbach G, Spitzer E, et al. VLA-4 and VCAM-1 are the principal adhesion molecules involved in the interaction between blast colony-forming cells and bone marrow stromal cells. Br J Haematol. 1995; 91:275–284. PMID: 8547062.
Article
12. Lichterfeld M, Martin S, Burkly L, Haas R, Kronenwett R. Mobilization of CD34+ haematopoietic stem cells is associated with a functional inactivation of the integrin very late antigen 4. Br J Haematol. 2000; 110:71–81. PMID: 10930981.
13. Lévesque JP, Hendy J, Takamatsu Y, Simmons PJ, Bendall LJ. Disruption of the CXCR4/CXCL12 chemotactic interaction during hematopoietic stem cell mobilization induced by GCSF or cyclophosphamide. J Clin Invest. 2003; 111:187–196. PMID: 12531874.
Article
14. Devine SM, Flomenberg N, Vesole DH, et al. Rapid mobilization of CD34+ cells following administration of the CXCR4 antagonist AMD3100 to patients with multiple myeloma and non-Hodgkin's lymphoma. J Clin Oncol. 2004; 22:1095–1102. PMID: 15020611.
Article
15. Girbl T, Lunzer V, Greil R, Namberger K, Hartmann TN. The CXCR4 and adhesion molecule expression of CD34+ hematopoietic cells mobilized by “on-demand” addition of plerixafor to granulocyte-colony-stimulating factor. Transfusion. 2014; 54:2325–2335. PMID: 24673458.
16. Avigdor A, Goichberg P, Shivtiel S, et al. CD44 and hyaluronic acid cooperate with SDF-1 in the trafficking of human CD34+ stem/progenitor cells to bone marrow. Blood. 2004; 103:2981–2989. PMID: 15070674.
Article
17. Szmigielska-Kaplon A, Szemraj J, Hamara K, et al. Polymorphism of CD44 influences the efficacy of CD34(+) cells mobilization in patients with hematological malignancies. Biol Blood Marrow Transplant. 2014; 20:986–991. PMID: 24680978.
Article
18. Bojko P, Pawloski D, Stellberg W, Schröder JK, Seeber S. Flt3 ligand and thrombopoietin serum levels during peripheral blood stem cell mobilization with chemotherapy and recombinant human glycosylated granulocyte colony-stimulating factor (rhu-G-CSF, lenograstim) and after high-dose chemotherapy. Ann Hematol. 2002; 81:522–528. PMID: 12373354.
19. Papayannopoulou T, Nakamoto B, Andrews RG, Lyman SD, Lee MY. In vivo effects of Flt3/Flk2 ligand on mobilization of hematopoietic progenitors in primates and potent synergistic enhancement with granulocyte colony-stimulating factor. Blood. 1997; 90:620–629. PMID: 9226162.
Article
20. He S, Chu J, Vasu S, et al. FLT3L and plerixafor combination increases hematopoietic stem cell mobilization and leads to improved transplantation outcome. Biol Blood Marrow Transplant. 2014; 20:309–313. PMID: 24365795.
Article
21. Zubair AC, Rymer R, Young J, et al. Multiple myeloma patients receiving large volume leukapheresis efficiently yield enough CD34+ cells to allow double transplants. J Clin Apher. 2009; 24:6–11. PMID: 19156756.
Article
22. Sutherland DR, Anderson L, Keeney M, Nayar R, Chin-Yee I. The ISHAGE guidelines for CD34+ cell determination by flow cytometry. International Society of Hematotherapy and Graft Engineering. J Hematother. 1996; 5:213–226. PMID: 8817388.
23. Petty JM, Lenox CC, Weiss DJ, Poynter ME, Suratt BT. Crosstalk between CXCR4/stromal derived factor-1 and VLA-4/VCAM-1 pathways regulates neutrophil retention in the bone marrow. J Immunol. 2009; 182:604–612. PMID: 19109194.
Article
24. Südhoff T, Söhngen D. Circulating endothelial adhesion molecules (sE-selectin, sVCAM-1 and sICAM-1) during rHuG-CSF-stimulated stem cell mobilization. J Hematother Stem Cell Res. 2002; 11:147–151. PMID: 11847011.
Article
25. Papayannopoulou T, Priestley GV, Nakamoto B, Zafiropoulos V, Scott LM. Molecular pathways in bone marrow homing: dominant role of alpha(4)beta(1) over beta(2)-integrins and selectins. Blood. 2001; 98:2403–2411. PMID: 11588037.
26. Lévesque JP, Takamatsu Y, Nilsson SK, Haylock DN, Simmons PJ. Vascular cell adhesion molecule-1 (CD106) is cleaved by neutrophil proteases in the bone marrow following hematopoietic progenitor cell mobilization by granulocyte colony-stimulating factor. Blood. 2001; 98:1289. PMID: 11520773.
27. Velders GA, Pruijt JF, Verzaal P, et al. Enhancement of G-CSF-induced stem cell mobilization by antibodies against the beta 2 integrins LFA-1 and Mac-1. Blood. 2002; 100:327–333. PMID: 12070044.
28. Gunji Y, Nakamura M, Hagiwara T, et al. Expression and function of adhesion molecules on human hematopoietic stem cells: CD34+ LFA-1- cells are more primitive than CD34+ LFA-1+ cells. Blood. 1992; 80:429–436. PMID: 1378320.
Article
29. Zöller M. CD44, hyaluronan, the hematopoietic stem cell, and leukemia-initiating cells. Front Immunol. 2015; 6:235. PMID: 26074915.
Article
30. Lapidot T, Petit I. Current understanding of stem cell mobilization: the roles of chemokines, proteolytic enzymes, adhesion molecules, cytokines, and stromal cells. Exp Hematol. 2002; 30:973–981. PMID: 12225788.
31. Lee S, Im SA, Yoo ES, et al. Mobilization kinetics of CD34(+) cells in association with modulation of CD44 and CD31 expression during continuous intravenous administration of G-CSF in normal donors. Stem Cells. 2000; 18:281–286. PMID: 10924094.
32. Fukuda S, Broxmeyer HE, Pelus LM. Flt3 ligand and the Flt3 receptor regulate hematopoietic cell migration by modulating the SDF-1alpha(CXCL12)/CXCR4 axis. Blood. 2005; 105:3117–3126. PMID: 15618475.
33. He S, Chu J, Vasu S, et al. FLT3L and plerixafor combination increases hematopoietic stem cell mobilization and leads to improved transplantation outcome. Biol Blood Marrow Transplant. 2014; 20:309–313. PMID: 24365795.
Article
34. Anandasabapathy N, Hurley A, Breton G, et al. A phase 1 trial of the hematopoietic growth factor CDX301 (rhuFlt3L) in healthy volunteers. Biol Blood Marrow Transplant. 2013; 19(Suppl):S112. (BMT Tandem Meetings Abstracts).
Article
35. Cheng M, Qin G. Progenitor cell mobilization and recruitment: SDF-1, CXCR4, α4-integrin, and c-kit. Prog Mol Biol Transl Sci. 2012; 111:243–264. PMID: 22917234.
Article
36. Carion A, Benboubker L, Hérault O, et al. Stromal-derived factor 1 and matrix metalloproteinase 9 levels in bone marrow and peripheral blood of patients mobilized by granulocyte colony-stimulating factor and chemotherapy. Relationship with mobilizing capacity of haematopoietic progenitor cells. Br J Haematol. 2003; 122:918–926. PMID: 12956762.
Article
37. Dar A, Kollet O, Lapidot T. Mutual, reciprocal SDF-1/CXCR4 interactions between hematopoietic and bone marrow stromal cells regulate human stem cell migration and development in NOD/SCID chimeric mice. Exp Hematol. 2006; 34:967–975. PMID: 16863903.
Article
38. Donahue RE, Jin P, Bonifacino AC, et al. Plerixafor (AMD3100) and granulocyte colony-stimulating factor (G-CSF) mobilize different CD34+ cell populations based on global gene and microRNA expression signatures. Blood. 2009; 114:2530–2541. PMID: 19602709.
Article
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