Quantitative events determine the differentiation and function of helper T cells


In recent years, numerous qualitative discoveries have been made in immunology research. However, the effect of quantitative events, long recognized as the driving factors for determinism in developmental biology, that dictate the quality of the immune response elicited to an antigen in concert with microbial products still requires serious attention. Here we discuss how the often-neglected issue of quantification affects the specification, differentiation and commitment of helper T cells. As reductionist in vitro approaches have been instrumental in the elucidation of the factors determining the development of helper T cells, in this perspective we highlight the need for the standardization of protocols, also fundamental for the comparison of immune responses in mice and humans. Improving understanding of how these in vitro quantitative events translate to immune responses in vivo, which can be studied in mouse models, is of importance in obtaining information on immune responses in humans, thus empowering translational research.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Plasticity and commitment of helper T cells.
Figure 2: Several factors determine the absolute amount of helper T cell cytokines produced in response to infection.


  1. 1

    Sher, A. & Coffman, R.L. Regulation of immunity to parasites by T cells and T cell-derived cytokines. Annu. Rev. Immunol. 10, 385–409 (1992).

  2. 2

    Constant, S.L. & Bottomly, K. Induction of Th1 and Th2 CD4+ T cell responses: the alternative approaches. Annu. Rev. Immunol. 15, 297–322 (1997).

  3. 3

    O'Garra, A. Cytokines induce the development of functionally heterogeneous T helper cell subsets. Immunity 8, 275–283 (1998).

  4. 4

    Glimcher, L.H. & Murphy, K.M. Lineage commitment in the immune system: the T helper lymphocyte grows up. Genes Dev. 14, 1693–1711 (2000).

  5. 5

    Casanova, J.L. & Abel, L. Genetic dissection of immunity to mycobacteria: the human model. Annu. Rev. Immunol. 20, 581–620 (2002).

  6. 6

    Cooper, A.M. Cell-mediated immune responses in tuberculosis. Annu. Rev. Immunol. 27, 393–422 (2009).

  7. 7

    Zhu, J., Yamane, H. & Paul, W.E. Differentiation of effector CD4 T cell populations. Annu. Rev. Immunol. 28, 445–489 (2010).

  8. 8

    Kolls, J.K. & Khader, S.A. The role of Th17 cytokines in primary mucosal immunity. Cytokine Growth Factor Rev. 21, 443–448 (2010).

  9. 9

    Littman, D.R. & Rudensky, A.Y. Th17 and regulatory T cells in mediating and restraining inflammation. Cell 140, 845–858 (2010).

  10. 10

    Stockinger, B. & Veldhoen, M. Differentiation and function of Th17 T cells. Curr. Opin. Immunol. 19, 281–286 (2007).

  11. 11

    Tato, C.M. & Cua, D.J. Reconciling id, ego, and superego within interleukin-23. Immunol. Rev. 226, 103–111 (2008).

  12. 12

    Duhen, T., Geiger, R., Jarrossay, D., Lanzavecchia, A. & Sallusto, F. Production of interleukin 22 but not interleukin 17 by a subset of human skin-homing memory T cells. Nat. Immunol. 10, 857–863 (2009).

  13. 13

    Trifari, S., Kaplan, C.D., Tran, E.H., Crellin, N.K. & Spits, H. Identification of a human helper T cell population that has abundant production of interleukin 22 and is distinct from TH-17, TH1 and TH2 cells. Nat. Immunol. 10, 864–871 (2009).

  14. 14

    Trifari, S. & Spits, H. IL-22-producing CD4+ T cells: middle-men between the immune system and its environment. Eur. J. Immunol. 40, 2369–2371 (2010).

  15. 15

    de Jong, A. et al. CD1a-autoreactive T cells are a normal component of the human αβ T cell repertoire. Nat. Immunol. 11, 1102–1109 (2010).

  16. 16

    Coombes, J.L. et al. A functionally specialized population of mucosal CD103+ DCs induces Foxp3+ regulatory T cells via a TGF-β and retinoic acid-dependent mechanism. J. Exp. Med. 204, 1757–1764 (2007).

  17. 17

    Mucida, D. et al. Retinoic acid can directly promote TGF-β-mediated Foxp3+ Treg cell conversion of naive T cells. Immunity 30, 471–472 (2009).

  18. 18

    Sun, C.M. et al. Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 Treg cells via retinoic acid. J. Exp. Med. 204, 1775–1785 (2007).

  19. 19

    Cruz, A. et al. Pathological role of interleukin 17 in mice subjected to repeated BCG vaccination after infection with Mycobacterium tuberculosis. J. Exp. Med. 207, 1609–1616 (2010).

  20. 20

    O'Shea, J.J. & Paul, W.E. Mechanisms underlying lineage commitment and plasticity of helper CD4+ T cells. Science (New York, N. Y 327, 1098–1102 (2010).

  21. 21

    Reinhardt, R.L., Khoruts, A., Merica, R., Zell, T. & Jenkins, M.K. Visualizing the generation of memory CD4 T cells in the whole body. Nature 410, 101–105 (2001).

  22. 22

    Fazilleau, N., McHeyzer-Williams, L.J., Rosen, H. & McHeyzer-Williams, M.G. The function of follicular helper T cells is regulated by the strength of T cell antigen receptor binding. Nat. Immunol. 10, 375–384 (2009).

  23. 23

    Reinhardt, R.L., Liang, H.E. & Locksley, R.M. Cytokine-secreting follicular T cells shape the antibody repertoire. Nat. Immunol. 10, 385–393 (2009).

  24. 24

    Johnston, R.J. et al. Bcl6 and Blimp-1 are reciprocal and antagonistic regulators of T follicular helper cell differentiation. Science 325, 1006–1010 (2009).

  25. 25

    Vinuesa, C.G., Linterman, M.A., Goodnow, C.C. & Randall, K.L. T cells and follicular dendritic cells in germinal center B-cell formation and selection. Immunol. Rev. 237, 72–89 (2010).

  26. 26

    Murphy, E. et al. Reversibility of T helper 1 and 2 populations is lost after long-term stimulation. J. Exp. Med. 183, 901–913 (1996).

  27. 27

    Mosmann, T.R., Cherwinski, H., Bond, M.W., Giedlin, M.A. & Coffman, R.L. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J. Immunol. 136, 2348–2357 (1986).

  28. 28

    Lee, G.R., Kim, S.T., Spilianakis, C.G., Fields, P.E. & Flavell, R.A. T helper cell differentiation: regulation by cis elements and epigenetics. Immunity 24, 369–379 (2006).

  29. 29

    Wohlfert, E. & Belkaid, Y. Plasticity of Treg at infected sites. Mucosal Immunol. 3, 213–215 (2010).

  30. 30

    Locksley, R.M. Nine lives: plasticity among T helper cell subsets. J. Exp. Med. 206, 1643–1646 (2009).

  31. 31

    Murphy, K.M. & Stockinger, B. Effector T cell plasticity: flexibility in the face of changing circumstances. Nat. Immunol. 11, 674–680 (2010).

  32. 32

    Fazilleau, N., McHeyzer-Williams, L.J. & McHeyzer-Williams, M.G. Local development of effector and memory T helper cells. Curr. Opin. Immunol. 19, 259–267 (2007).

  33. 33

    Malherbe, L., Mark, L., Fazilleau, N., McHeyzer-Williams, L.J. & McHeyzer-Williams, M.G. Vaccine adjuvants alter TCR-based selection thresholds. Immunity 28, 698–709 (2008).

  34. 34

    O'Garra, A. & Murphy, K.M. From IL-10 to IL-12: how pathogens and their products stimulate APCs to induce TH1 development. Nat. Immunol. 10, 929–932 (2009).

  35. 35

    Constant, S., Pfeiffer, C., Woodard, A., Pasqualini, T. & Bottomly, K. Extent of T cell receptor ligation can determine the functional differentiation of naive CD4+ T cells. J. Exp. Med. 182, 1591–1596 (1995).

  36. 36

    Hosken, N.A., Shibuya, K., Heath, A.W., Murphy, K.M. & O'Garra, A. The effect of antigen dose on CD4+ T helper cell phenotype development in a T cell receptor-αβ-transgenic model. J. Exp. Med. 182, 1579–1584 (1995).

  37. 37

    Iezzi, G. et al. CD40–CD40L cross-talk integrates strong antigenic signals and microbial stimuli to induce development of IL-17-producing CD4+ T cells. Proc. Natl. Acad. Sci. USA 106, 876–881 (2009).

  38. 38

    Langenkamp, A., Messi, M., Lanzavecchia, A. & Sallusto, F. Kinetics of dendritic cell activation: impact on priming of TH1, TH2 and nonpolarized T cells. Nat. Immunol. 1, 311–316 (2000).

  39. 39

    Nembrini, C., Abel, B., Kopf, M. & Marsland, B.J. Strong TCR signaling, TLR ligands, and cytokine redundancies ensure robust development of type 1 effector T cells. J. Immunol. 176, 7180–7188 (2006).

  40. 40

    Ruedl, C., Bachmann, M.F. & Kopf, M. The antigen dose determines T helper subset development by regulation of CD40 ligand. Eur. J. Immunol. 30, 2056–2064 (2000).

  41. 41

    Valitutti, S., Muller, S., Cella, M., Padovan, E. & Lanzavecchia, A. Serial triggering of many T-cell receptors by a few peptide-MHC complexes. Nature 375, 148–151 (1995).

  42. 42

    Viola, A., Schroeder, S., Sakakibara, Y. & Lanzavecchia, A. T lymphocyte costimulation mediated by reorganization of membrane microdomains. Science 283, 680–682 (1999).

  43. 43

    Viola, A. et al. Quantitative contribution of CD4 and CD8 to T cell antigen receptor serial triggering. J. Exp. Med. 186, 1775–1779 (1997).

  44. 44

    Gett, A.V., Sallusto, F., Lanzavecchia, A. & Geginat, J. T cell fitness determined by signal strength. Nat. Immunol. 4, 355–360 (2003).

  45. 45

    Iezzi, G., Karjalainen, K. & Lanzavecchia, A. The duration of antigenic stimulation determines the fate of naive and effector T cells. Immunity 8, 89–95 (1998).

  46. 46

    Iezzi, G., Scotet, E., Scheidegger, D. & Lanzavecchia, A. The interplay between the duration of TCR and cytokine signaling determines T cell polarization. Eur. J. Immunol. 29, 4092–4101 (1999).

  47. 47

    Lee, K.H. et al. The immunological synapse balances T cell receptor signaling and degradation. Science 302, 1218–1222 (2003).

  48. 48

    Miller, M.J., Safrina, O., Parker, I. & Cahalan, M.D. Imaging the single cell dynamics of CD4+ T cell activation by dendritic cells in lymph nodes. J. Exp. Med. 200, 847–856 (2004).

  49. 49

    Paulos, C.M. et al. The inducible costimulator (ICOS) is critical for the development of human TH17 cells. Sci. Transl. Med. 2, 55ra78 (2010).

  50. 50

    Bauquet, A.T. et al. The costimulatory molecule ICOS regulates the expression of c-Maf and IL-21 in the development of follicular T helper cells and TH-17 cells. Nat. Immunol. 10, 167–175 (2009).

  51. 51

    Saraiva, M. & O'Garra, A. The regulation of IL-10 production by immune cells. Nat. Rev. Immunol. 10, 167–175 (2009).

  52. 52

    Tao, X., Constant, S., Jorritsma, P. & Bottomly, K. Strength of TCR signal determines the costimulatory requirements for Th1 and Th2 CD4+ T cell differentiation. J. Immunol. 159, 5956–5963 (1997).

  53. 53

    Jorritsma, P.J., Brogdon, J.L. & Bottomly, K. Role of TCR-induced extracellular signal-regulated kinase activation in the regulation of early IL-4 expression in naive CD4+ T cells. J. Immunol. 170, 2427–2434 (2003).

  54. 54

    Yamane, H., Zhu, J. & Paul, W.E. Independent roles for IL-2 and GATA-3 in stimulating naive CD4+ T cells to generate a Th2-inducing cytokine environment. J. Exp. Med. 202, 793–804 (2005).

  55. 55

    Pfeiffer, C. et al. Altered peptide ligands can control CD4 T lymphocyte differentiation in vivo. J. Exp. Med. 181, 1569–1574 (1995).

  56. 56

    Aguado, E. et al. Induction of T helper type 2 immunity by a point mutation in the LAT adaptor. Science 296, 2036–2040 (2002).

  57. 57

    Aguilar-Pimentel, J.A. et al. Specific CD8 T cells in IgE-mediated allergy correlate with allergen dose and allergic phenotype. Am. J. Respir. Crit. Care Med. 181, 7–16 (2010).

  58. 58

    Jakob, T. et al. Novel mouse mutants with primary cellular immunodeficiencies generated by genome-wide mutagenesis. J. Allergy Clin. Immunol. 121, 179–184 (2008).

  59. 59

    Jun, J.E. et al. Identifying the MAGUK protein Carma-1 as a central regulator of humoral immune responses and atopy by genome-wide mouse mutagenesis. Immunity 18, 751–762 (2003).

  60. 60

    Malissen, B., Aguado, E. & Malissen, M. Role of the LAT adaptor in T-cell development and Th2 differentiation. Adv. Immunol. 87, 1–25 (2005).

  61. 61

    Siggs, O.M. et al. Opposing functions of the T cell receptor kinase ZAP-70 in immunity and tolerance differentially titrate in response to nucleotide substitutions. Immunity 27, 912–926 (2007).

  62. 62

    Sommers, C.L. et al. A LAT mutation that inhibits T cell development yet induces lymphoproliferation. Science 296, 2040–2043 (2002).

  63. 63

    Steinfelder, S. et al. The major component in schistosome eggs responsible for conditioning dendritic cells for Th2 polarization is a T2 ribonuclease (omega-1). J. Exp. Med. 206, 1681–1690 (2009).

  64. 64

    Milner, J.D., Fazilleau, N., McHeyzer-Williams, M. & Paul, W. Cutting edge: lack of high affinity competition for peptide in polyclonal CD4+ responses unmasks IL-4 production. J. Immunol. 184, 6569–6573 (2010).

  65. 65

    Eisenbarth, S.C. et al. Lipopolysaccharide-enhanced, toll-like receptor 4-dependent T helper cell type 2 responses to inhaled antigen. J. Exp. Med. 196, 1645–1651 (2002).

  66. 66

    Hammad, H. et al. House dust mite allergen induces asthma via Toll-like receptor 4 triggering of airway structural cells. Nat. Med. 15, 410–416 (2009).

  67. 67

    Hill, J.A. et al. Retinoic acid enhances Foxp3 induction indirectly by relieving inhibition from CD4+CD44hi cells. Immunity 29, 758–770 (2008).

  68. 68

    Nolting, J. et al. Retinoic acid can enhance conversion of naive into regulatory T cells independently of secreted cytokines. J. Exp. Med. 206, 2131–2139 (2009).

  69. 69

    Kretschmer, K. et al. Inducing and expanding regulatory T cell populations by foreign antigen. Nat. Immunol. 6, 1219–1227 (2005).

  70. 70

    Gabryšová, L. et al. Integrated T cell receptor and costimulatory signals determine TGFβ-dependent differentiation and maintenance of Foxp3+ regulatory T cells. Eur. J. Immunol. published online, doi:10.1002/eji.201041073 (24 February 2011).

  71. 71

    Oliveira, V.G., Caridade, M., Paiva, R.S., Demengeot, J. & Graca, L. Sub-optimal CD4 T cell activation triggers autonomous TGF-β-dependent conversion to Foxp3+ regulatory T cells. Eur. J. Immunol. published online, doi:10.1002/eji.201040896 (24 February 2011).

  72. 72

    Turner, M.S., Kane, L.P. & Morel, P.A. Dominant role of antigen dose in CD4+Foxp3+ regulatory T cell induction and expansion. J. Immunol. 183, 4895–4903 (2009).

  73. 73

    Battaglia, M., Stabilini, A. & Roncarolo, M.G. Rapamycin selectively expands CD4+CD25+FoxP3+ regulatory T cells. Blood 105, 4743–4748 (2005).

  74. 74

    Sauer, S. et al. T cell receptor signaling controls Foxp3 expression via PI3K, Akt, and mTOR. Proc. Natl. Acad. Sci. USA 105, 7797–7802 (2008).

  75. 75

    Wing, K. & Sakaguchi, S. Regulatory T cells exert checks and balances on self tolerance and autoimmunity. Nat. Immunol. 11, 7–13 (2010).

  76. 76

    Gabrysova, L. et al. Negative feedback control of the autoimmune response through antigen-induced differentiation of IL-10-secreting Th1 cells. J. Exp. Med. 206, 1755–1767 (2009).

  77. 77

    Gabrysova, L. & Wraith, D.C. Antigenic strength controls the generation of antigen-specific IL-10-secreting T regulatory cells. Eur. J. Immunol. 40, 1386–1395 (2010).

  78. 78

    O'Garra, A. & Vieira, P. Regulatory T cells and mechanisms of immune system control. Nat. Med. 10, 801–805 (2004).

  79. 79

    O'Garra, A. & Vieira, P. TH1 cells control themselves by producing interleukin-10. Nat. Rev. Immunol. 7, 425–428 (2007).

  80. 80

    Trinchieri, G. Interleukin-10 production by effector T cells: Th1 cells show self control. J. Exp. Med. 204, 239–243 (2007).

  81. 81

    Saraiva, M. et al. Interleukin-10 production by Th1 cells requires interleukin-12-induced STAT4 transcription factor and ERK MAP kinase activation by high antigen dose. Immunity 31, 209–219 (2009).

  82. 82

    Dresser, D.W. Specific inhibition of antibody production. II. Paralysis induced in adult mice by small quantities of protein antigen. Immunology 5, 378–388 (1962).

  83. 83

    Dresser, D.W. Specific inhibition of antibody production. I. Protein-over loading paralysis. Immunology 5, 161–168 (1962).

  84. 84

    Mitchison, N.A. Induction of immunological paralysis in two zones of dosage. Proc. R. Soc. Lond. B 161, 275–292 (1964).

  85. 85

    Akdis, C.A., Joss, A., Akdis, M. & Blaser, K. Mechanism of IL-10-induced T cell inactivation in allergic inflammation and normal response to allergens. Int. Arch. Allergy Immunol. 124, 180–182 (2001).

  86. 86

    Meiler, F. et al. In vivo switch to IL-10-secreting T regulatory cells in high dose allergen exposure. J. Exp. Med. 205, 2887–2898 (2008).

  87. 87

    Parish, C.R. & Liew, F.Y. Immune response to chemically modified flagellin. 3. Enhanced cell-mediated immunity during high and low zone antibody tolerance to flagellin. J. Exp. Med. 135, 298–311 (1972).

  88. 88

    Liew, F.Y. TH1 and TH2 cells: a historical perspective. Nat. Rev. Immunol. 2, 55–60 (2002).

  89. 89

    Mosmann, T.R. & Coffman, R.L. Heterogeneity of cytokine secretion patterns and functions of helper T cells. Adv. Immunol. 46, 111–147 (1989).

  90. 90

    Redford, P.S. et al. Enhanced protection to Mycobacterium tuberculosis infection in IL-10-deficient mice is accompanied by early and enhanced Th1 responses in the lung. Eur. J. Immunol. 40, 2200–2210 (2010).

  91. 91

    Anderson, P. Post-transcriptional control of cytokine production. Nat. Immunol. 9, 353–359 (2008).

  92. 92

    Pascual, V., Chaussabel, D. & Banchereau, J. A genomic approach to human autoimmune diseases. Annu. Rev. Immunol. 28, 535–571 (2010).

  93. 93

    Pulendran, B., Li, S. & Nakaya, H.I. Systems vaccinology. Immunity 33, 516–529 (2010).

  94. 94

    Green, J.B. & Smith, J.C. Growth factors as morphogens: do gradients and thresholds establish body plan? Trends Genet. 7, 245–250 (1991).

  95. 95

    Ansel, K.M., Djuretic, I., Tanasa, B. & Rao, A. Regulation of Th2 differentiation and Il4 locus accessibility. Annu. Rev. Immunol. 24, 607–656 (2006).

  96. 96

    Messi, M. et al. Memory and flexibility of cytokine gene expression as separable properties of human TH1 and TH2 lymphocytes. Nat. Immunol. 4, 78–86 (2003).

  97. 97

    Agarwal, S. & Rao, A. Modulation of chromatin structure regulates cytokine gene expression during T cell differentiation. Immunity 9, 765–775 (1998).

  98. 98

    Veldhoen, M., Hirota, K., Christensen, J., O'Garra, A. & Stockinger, B. Natural agonists for aryl hydrocarbon receptor in culture medium are essential for optimal differentiation of Th17 T cells. J. Exp. Med. 206, 43–49 (2009).

  99. 99

    Ghoreschi, K. et al. Generation of pathogenic TH17 cells in the absence of TGF-β signalling. Nature 467, 967–971 (2010).

  100. 100

    Yang, X. et al. Opposing regulation of the locus encoding IL-17 through direct, reciprocal actions of STAT3 and STAT5. Nat. Immunol. 12, 247–254 (2011).

Download references


We thank J. Langhorne, A. Potocnik, G. Kassiotis and A. Howes for review of the manuscript and comments, and P. Redford for compiling Figure 2. Supported by the Medical Research Council UK (A.O.G. and L.G.).

Author information



Corresponding author

Correspondence to Hergen Spits.

Ethics declarations

Competing interests

H.S. works one day per week for AIMM Therapeutics.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

O'Garra, A., Gabryšová, L. & Spits, H. Quantitative events determine the differentiation and function of helper T cells. Nat Immunol 12, 288–294 (2011). https://doi.org/10.1038/ni.2003

Download citation

Further reading