IMMUNOLOGY TODAY

The expanding universe of T-cell

subsets: Thl, Th2 and more
Tim R. Mosmann and Subash Sad

he T helper 1 (Thl) and Th2 How many discrete cytokine

patterns of cytokine produc- patterns are expressed by C
tion were originally described T cells?
among mouse CD4’ T-cell Although there are many well-documented
clone@ and later among human T cells’. Thl and Th2 responses, as summarized in
Mouse Thl cells produce interleukin 2 (IL-2), Table 2, these are not the only cytokine
interferon y (IFN-y) and lymphotoxin (LT), patterns possible: T cells expressing cyto-
whereas Th2 cells produce IL-4, IL-5, IL-6, kines of both patterns have been called ThO
IL-9, IL-10 and IL-13. Human Thl and Th2 cells”; ceils producing high amounts of
cells produce similar patterns, although the transforming growth factor 8 (TGF-B) have
synthesis of IL-2, IL-6, IL-10 and IL-13 is not been termed Th3 (Ref. 11); and additional
as tightly restricted to a single subset as phenotypes have been described among
in mouse T cells. Several other proteins are long-term clones. At the single-cell level
secreted both by Thl and Th2 cells, includ- among normal cells in short-term culture,
ing IL-3, tumor necrosis factor a (TNF-o), IL-4 and IL-5 are often co-expressed, IL-4
granulocyte-macrophage colony-stimulating factr r (GM-CSF), and IFN-y are normally exclusively expressed, and IFN-y and IL-10
[Metlenkephalin and members of the chemokine (CR) families are co-expressed in some cells’2-‘5.There may even be heterogeneity
(Table 1). As the full range of cytokines has not been determined in within the Thl and Th2 subsets if the large quantitative variations
many studies, the terms Thl or Th2 (or type 1 or type 2) will be used in cytokine secretion that are observed between clones in vitro also
here to refer to either the full cytokine patterns defined by mouse occur in normal cells. Possible interpretations of subset complexity
T-cell clones, or to responses dominated by IFN-7/or IL-4, respectively. range from a simple Thl /Th2 (and ThO/Th3?) model with quanti-
It should be noted that this may underestimate the full complexity tative differences at different development stages, to a model in
of some responses. which there are no discrete subsets but rather a continuum of dif-
The functions of Thl and Th2 cells correlate well with their dis- ferent combinations of cytokine secretioni6. In some systems, clones
tinctive cytokines. Thl cells are involved in cell-mediated inflam- show expression patterns of Thl and Th2 cytokines that fit a
matory reactions: several Thl cytokines activate cytotoxic and random distribution”j. However, analysis of data from an earlier
inflammatory functions4; Thl clones induce delayed-type hyper- study reveals that short-term allospecific clones from unimmunized
sensitivity (DTH) reactions; and IFNy is commonly expressed at mice, or mice infected with Nippostror~gyfus brnsilicnsis, showed a
sites of DTH reactions5,h. Some B-cell help can be provided by Thl striking inverse correlation of IFN-y and IL-4 synthesis, whereas
cells but, at higher Thl-cell numbers, this can become suppression7s8. IL-4 and IL-5 synthesis showed a positive correlation” (Fig. 1).
Th2 cytokines encourage antibody production, particularly IgE re- Patterns of single-cell cytokine expression by cells in short-term cul-
sponses”, and also enhance eosinophil proliferation and function. ture’Z-‘5 also indicate that, while there are clearly more patterns
Accordingly, Th2 cytokines are commoniy found in association with than the extreme Thl/Th2 pattern, cytokine expression is normally
strong antibody and allergic responses. nonrandom.
The characteristic cytokine products of Thl and Th2 cells are mutu- If cytokine expression is nonrandom, how many distinct pheno-
ally inhibitory for the differentiation and effector functions of the recip- types of T cdl can exist? Minor or quantitative differences in cyto-
rocal phenotype. Thus, IFN-7 selectively inhibits proliferation of Th2 kine expression may represent distinct phenotypes, developmental
cells4, and IL-10 inhibits cytokine synthesis by Thl cells9. This cross- stages, or transient responses to stimulation conditions. Cytokine
regulation may partly explain the strong biases towards Thl or Th2 patterns can also be changed by strong stimuli or other cytokines:
responses during many infections in mice and humans (Table 2). In for instance, IL-2 and IL-12 (Ref. 18) enhance 1lW-y synthesis,
severaI cases, alteration of these patterns by cytokine or anti-cytokine whereas IL-10 inhibits Thl cytokine synthesis9. This might allow
reagents reverses host resistance or susceptibility to infection. Thus, short-term modulation of the cytokine response without permanent
there is ample evidence that these cytokine patterns are important in alteration of the T-cell phenotypes. Other phenotypes might be
mediating resistance to several infectious agents (reviewed in Ref. 10). transient stages along a differentiation pathway, such as the ThOA
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MARCH -
 IMMUNOLOGY TODAY

(IL-2, IL-41 and ThOB (IL-2, IL-4, IFN-V) pat-

le ems
tern, which could undergo further dif-
ferentiation”. Single-cell cytokine-staining
Cell type Thl Common Th2 Other
methods, now successfully developed in
several laboratories, will be invaluable in re- Thl IL-2, IFN-y, LT IL-3,GM, TNF,CK
solving the contributions of these different Tel IL-2,IFNq, LT GM. TNF,CK
types of diversity.
Regardless of whether the variation in T-y&I IFN-y
T-cell cytokine synthesis represents a contin- NK IFN-y
uum or discrete subsets, there is no doubt
that many T-cell clones and in niuo immune ThO 11-2,IFN-y. LT 11-3,GM, TNF, CK IL-4, -5, -6, -9, - IO
responses show a dramatic dichotomy be-
tween IL-2/IFN-y/LT and IL-4/IL-5/IL-10
Th2 IL-3, GM, TNF, CK lL-4,-5, -6, -9, -IO
responses. Thus, the Thl-Th2 dichotomy
remains an important functional division in Tc2 IL-3,GM, TNF,CK 11-4,-5, -6, -9, -IO
the immune system. T$&2 IL-4

Mast cell IL-3,TNF 11-4,IL-IO

ifferentiation of Th I and Th2 CD3+CD4+NK I. I+ IL-4
cells from a common II.-2-secreting
B cell 11-4,IL-IO
precursor
Thl and Th2 patterns of cytokine secretion
correspond to activated effector pheno- Macrophage TNF, CK IL-6, IL-IO IL-I. IL-12
types generated during an immune re- Keratinocyte GM, TNF IL-IO IL-I
sponse. They do not exist among naive -
T cells, or possibly even among long-term Abbreviations:CK, chemokine; GM,granulocyt~macrophage colony-stimulatingfactor; IFN?,
interfemn 7; IL-I, interldin 1; LT,lymphotoxin;MC,natural killer;TqS,T cell with yS T-cellreceptor;
memory cells. When first stimulated by
Tc,T cytotoxic;Th, T helper; TNF,tumor nemesis factor.
antigen on antigen-presenting cells (AI%),
naive CD4’ T cells principally or entirely
produce IL-2, and then differentiate into phenotypes that secrete Factors affecting Th I ITh2 &ffevetbtIation
other cytokines (Fig. 2). Thus, Thl and Th2 cells can both be The control of differentiation of uncommitted T-cell precursors
derived from a single precursor cell. Using TGF-8 and anti-IFN-y should be distinguished from the regulation of the resulting effcc-
to inhibit Th2 and Thl differentiation, it is possible to keep tor cells since some, but not all, of the cytokines influencing these
CD4+ T cells in a proliferating, IL-2-secreting state20. These cells two processes are the same. IL-4 stimulates differentiation into Th2
lose their original, naive surface-antigen phenotype fCD44’“, cells, whereas IFN--f, IL-12 and TGF-8 enhance Thl developmentr’.
CD45RBhi, L-selectin”‘) and express a phenotype reminiscent of However, the role of IFN-y may be variable in different experimen-
memory cells (CD44 hi, CD45RB’“, L-seIectin’“). Subsequent re- tal systems: Thl differentiation is inhibited in some systemsz0,23*2”
by
stimulation of such cells in the presence of IL-4 or TGF-l3 results in anti-IFN-y, but not in all?“.
the generation of Th2- or Thl-like cytokine-secretion patterns, In a primary response, antigen-specific T cells are present at very
respectively. low frequency (e.g. 1 in 105) and other cells may provide the early
The differentiation of effector Thl /Th2 phenotypes may proceed IFN-1 or IL-4 that influences T-cell differentiation. IL-12 is made by
through an intermediate state expressing multiple cytokines. This many cell types, but particularly by macrophages in response to
was demonstrated using a transgenic mouse expressing a herpes- certain microbial products. Natural killer (NK) cells synthesize EN-1
virus thymidine kinase (HSV-TK) gene under the control of the IL4 in response either to IL-12 (Ref. 18) or to antigen-mediated cross-
promoter. IL-4 promoter activity occurred during differentiation of linking of antibody bound to Fc receptorsz5. In mice, early IL-4 pro-
naive cells into either Thl or Th2 phenotypes, since gancyclovir duction may be contributed by mast cells, basophils and a recently
(which is toxic to cells expressing HSV-TK) prevented appearance discovered subpopulation of CQ3+CD4+NKl.l’ T cells that appear
of both cell types2i. Transient expression of endogenous IL-4 mRNA to be restricted by major histocompatibility complex (MHCl class I
could also be detected during Thl differentiation (R. Flavell, pers. molecules26. These cells have a restricted T-cell receptor (TCR) rep-
commun.). It is not yet clear whether IL-4 mRNA expression is ac- ertoire, unknown antigen specificity and produce large amounts of
companied by protein secretion, whether IFN-,y is expressed on the IL-4 within a few hours of polyclonal stimulation.
Th2 differentiation pathway, or whether the full ThO phenotype Although initial expectations were that different ARC types
k expressed as an obligate intermediate on the differentiation would selectively influence T-cell differentiation, it is now known
pathway. that dendritic cells,, macrophages and B cells are all capable of
IMMUNOLOGY TODAY

Table 2. Thl and Th2 responses

Azenfldisease Host Response Comments Refs

Leishmaniamajor M Thl cures Th I response in resistant mice, Th2 in susceptible: anti-IL-4 protects, 55
anti-IFN-y exacerbates; Th I cells protect, Th2 cells exacerbate

I_ denovoni H Thl cures? IL-4 higher in visceral (generalized) leishmaniasis, IFN-7 higher in cutaneous 56
(local) leishmaniasis

L bmsiliensis H Thl cures? Th I cytokines higher in localized, DTH reactions; mixed cytokine patterns, 57
but higher IL4 in chronic, destructive mucocutaneous leishmaniasis; IFN-y
increases with cure

Tryponosomo ctwzi M Th I protects Th I cells transfer protection 58

T: equiperdium M Th I protects Successful immunization associated with a Th I response 59

f/osmedium chabaudi M Thl and Th2 Th I response provides early NO-dependent defense, Th2 provides later 60
antibody-dependent defense
Helminths
Ttiuris muris M Th2 cures Th2 response in resistant mice, Th I in susceptible: anti-IFN-y protects, 61
IL4 protects, anti-IL4 exacerbates
Nippostmrtgyk~s M Th2 cures Th2 response associated with rejection; IFN-1 and IL-I 2 reduce protection; 62
bmsijiensis IL4 accelerates expulsion
Heligmesomoides M Th2 cures Sustained Th2 response associated with expulsion; IL4 accelerates expulsion 63
p419yNs
Bfugio majayi M Th2 protects Th2 response correlates with protection, Th I with lack of protection 64
Schistosomo monsoni M Th2 response Schistosome eggs induce Th2 response; Th2 response associated with 65
granulomas, reduced by IL- I2 or anti-IL-4 treatment
vlruaer
HIV H Th I-Th2 switch IL4 responses increased, IFN-v responses decreased, in HIV+ patients 66,67
progressing to AIDS; restored by IL- I2 or anti-IL-4 in vitro; controversial
data
Th I to ThO Larger number of Feel1 clones from HIV* patients make IL-4 and are 37
ThO clones
Th2 susceptible Th2 clones are more susceptible to HIV infection than Th I
Tc2 response Tc2 clones (Th2 cytokines, CDS*) isolated from subset of HIV+ patients, 37
but not controls
MAIDS Th2 response Th2 response (cytokines, IgE) associated with disease progression 68
Measles Th2 response Vaccination causes increased IL4 production and transient inhibition 69
of third-party DTH responses’
Vaccinia Th2 exacerbates Concurrent schistosome infection weakens the anti-vaccinia response; virus 70
persists in granulomas
HSV Th2 exacerbates HSV-specific Th2 clone exacerbates HSV=induced keratitis 71
Influenza virus Thl protects Th I, but not Th2, anti-influenza clones confer in vivoprotection 72
Sezary syndrome Th2 114, IL-5 and IL-IO, but not IFN-y, elevated in PBMCs and skin lesions: 73
(HTLV.1) eosinophilia, elevated IgE
Mycosis fungoides Thl IFN-y, but not IL-4, IL-S, IL- IO, present in skin lesions 73
(HTLV- I)

Fungi
Gll?dida Thl protects Th I and Th2 responses correlate with resistance and susceptibility; anti-lL4, 74
anti-IL= IO and soluble IL4R induce resistance
Bacteria
Th I response IFN-y. but not IL4 or IL-S, expressed in pleural fluid, correlating with 75
resistant response: IL- IO also expressed (by macrophages)

1996
 IMMUNOLOGY TODAY

BCG H Th I response
M. leprae H ThIIDTH, Th;Z/Ab IL-2 and IFN-y expressed in tuberculoid (DTH) leprosy lesions; lL4, IL-5 and
M Th2 protects
Borderellapert&s M Thl cells
Chlamydial infection M Thl protects
Lisreria menocytogenes M Th I response
IL-2 and IFN-?/ expressed at the site of tuberculin (DTH) reaction 6
5
IL- IO expressed in lepromatous (Ab) lesions
Th I clones specific for mycobacterial antigens isolated from tuberculoid 76
leprosy (DTH) patients
Th2 antigen-specific cells confer resistance; resistant mice produce more IL4, 77
resistance weakened by anti-IL4 susceptible C3H mice produce more IFN-y.
susceptibility reduced by anti-IFN-y
Th I response; Th I cells transfer protection 78
IFN-y produced during resistant response: anti-IFN-y exacerbates, IFN-v 79
protects
Th I response induced, anti-IFN-y exacerbates 29

Autoimmunity
Psoriasis vulgaris H Th I response IFN-y. but not IL-4, produced by T-cell clones from psoriasis sites, and 73,80
expressed in lesions
EAE M Thl causes disease Thl clones induce disease; IFNq, but not IL-4, expressed by T cells in brain; 51
anti-IL- I2 ameliorates disease
Th3 protects TGF-B-producing cells protect: anti-TGF-B reverses protection II
MS H Thl anti-MBP Fcell clones from MS patients are specific for MBP and are Th I 81
Experimental R Th I causes, Th2 IFN-y-producing T cells induce disease, IL4-producing cells 82
autoimmune prevents (induced by HgClJ prevent
uveoretinitis
IDDM M Thl? IFN-y and IL4 expression correlate with B-cell destruction or protection, 51
respectively
RA H Th I response Synovial tissue from RA patients produces IFN-1, but not IL4 83
ReA H Th I response Yenenia entemfytica-specificT cells (associated with ReA) are lh I 84

Allergy
Allergy H Th2 response IL4 and IL-5, but not IFN-y. detected in late-phase cutaneous reaction; 3, 85
allergen-specific clones show a strong bias towards the Th2 phenotype
Atopic asthma H Th2 response Th2 cytokines detected in bronchial lavage; eosinophilia 86
Vernal conjunctivitis H Th2 response Fcell clones from conjunctivitis produce more IL4 and less IFN-11 than 87
control cells
Omen& syndrome H Th2 response lmmunodeficiency accompanied by eosinophilia, high IgE levels, high IL4 and 88
11-5, and low IFN-y; IFNq produces improvement
Transplantationand pregnancy
Transplant rejection M Th I response Several cytokines upregulated in rejecting liver, particularly IFNq 89
R Th I response Rejection correlates with higher IFN-y and lower IL4 levels; anti-CD4 90
theapy induces nonrejection and the opposite cytokine pattern
H Th I response? IL-2 higher in strongly rejecting heart allografu; IL-4 and IL-IO higher in mild 91
rejection
Chronic GVHD M Th2 response Increased spontaneous and induced levels of 114, but not IFN--y 92

Acute GVHD M Th I response Elevated levels of IFN--y in response to ConA 93

Pregnancy H Th I suppressed Cell-mediated immunity inhibited in infections and autoimmunity 52

M Thl harms, Several cytokines produced spontaneously in placenta; IL-2, IFN-?I, TNF and 94
Th2 protects NK cells increase resorptions; IL- IO protects

Abbreviations: Ab, antibody; BCG,baciIIeCabnette-Gu&in; Con& concanavalinA; DTH,delayed-type hypersensitivity; EAE,experimental autoimmune
encephalomyelitie;GVHD,graft-versus-host disease; H, human; HIV,human immnnodeficiency virus; HSV,herpes simplex virus; HTLV-I,human T-cell
leukemia virus type 1; IDDM,inaulindependent diabetes melhtua; IPN?, interferon ‘y;IL-l, interleukin 1; IL-4R,IL-4receptor; LT,lymphotoxin;
M, mouse; MAIDS,mouse acquired imm~odeficiency syndrome; MBP,myelin basic protein; MS,multiple sclerosis; NK, natural kiier; NO, nitric oxide;
PBMC,peripheral blood mononudear cell; R, rat; RA, rheumatoid arthritis; ReA, reactive arthritis; Tc,T cytotoxic;TGF-B,transforming growth factor B;Th,
T helper; TNP,tumor necrosis factor.

MARCH I996
IMMUNOLOGY TODAY

Unimmunized

Nippos trongy/us

* . * A.”v
.
i
*
la +
0
n A
A
0

Fig. I. Nonrnndom
cytokine patterns of shot?-term allospecifk T-cell clones. Allospecific T-cell clones were isolated from rrnirnmunized or
Nippostrongylus-infected mice. After 56 (red triangle) and 70 (blue triangle) days of culture, the cells were stimulated with concurzavalinA, and
cytokines were measured in the suaernatnnts after 24 hours. These data indicate that K-4 rend K-5 levels slrow a positive correlation, whereas IL4 and
IFN-y show a negative correlation. Data in this figure are replotted from Ref, 17. Abbreviations: IFN-y, interferon y; E-4, interleukin 4.

inducing differentiation of Thl or Th2 cells in the presence of the matches the appropriate response to a particular pathogen. This is
appropriate cytokines. However, this does not exclude a role for dif- an important decision and we may expect precise and multiple
ferent APCs in the Thl-Th2 decision, as they may ultimately exert mechanisms to translate the properties of the pathogen into ryto-
selective influences depending on the cytokines they secrete. Further- kine or costimulatory signals that induce the appropriate type of
more, recent evidence suggests that costimulatory molecules on T cells and effector functions. For example, it has been suggested
APCs may selectiv:ly influence T-cell differentiation: antibodies that proteinases, required by multicellular organisms for tissue pen-
against B7.1 or 87.2 selectively inhibit the development of Thl and etration, induce an appropriate Th2 response against multicellular
Th2 responses, respectively, both in vitro and in vivo (reviewed in parasites 28.Furthermore, a Listeria product induces IL-12 synthesis,
Ref. 27). However, the contribution of B7.1 and B7.2 to T-cell differ- thus initiating a Thl response29.
entiation may be more complex than initially thought. Both B7.1
and B7.2 can induce Thl or Th2 differentiation, and their relative
contributions may be affected by antigen concentration. 87.2 appears Low and high antigen dose
to be the dominant costimulatory molecule during primary responses, Antigen dose may be a major regulator of the choice of effector
whereas B7.1, which is upregulated later in immune responses, may functions. Early in vivo studies showed that both low and high
be critical in maintaining primary and secondary immune responses. amounts of antigen primed for DTH, whereas moderate levels
stimulated antibody production”“. Similar effects occur in infectious
models, as shown by the induction of a healing, DTH response in
How is the Th I-Th2 choice made? BALB/c mice infected with very low numbers of Leishmania para-
Although several cytokines have well-characterized effects on T-cell sites, and a nonhealing, antibody response with higher dose@.
differentiation (Fig. 2), it is less clear how the immune system Recent experiments in vitro have also impbcated antigen dose as an
 1MMUNOLOGY TODAY

anti-IFN-y
important factor in the Thl-Th2 decision,
although the interpretation of the results is
complex. Increasing antigen dose may
switch the immune response from Thl to
Th2 or vice versa and, when a wide
range of antigen concentrations is used,
two switching events can occur (i.e.
Th2+Thl+Th2)32. However, results in U~VO~~
suggested a DTH+antibody*DTH pro-
gression. Once again, the effects of antigen
concentration on differentiation must be
distinguished from the regulation of mature
Thl and Th2 effector cells: fully differenti-
ated Thl cells are stimulated by moderate
but not high doses of antigen, whereas Th2
cells respond over a wider range of
concentrations33.
The consequences of ‘low’ or ‘high’ anti-
gen doses may depend on the soluble or
particulate nature of the antigen. At low
concentrations of a soluble antigenic pep-
tide, each AK should present a low density
of peptide-MHC complexes. By contrast, at
low doses of an infectious agent, or in the CD8+ precursors
presence of heterologous erythrocytes, al-
though relatively few APCs capture anti-
gen, each successful APC should present
substantial amounts because of the diges- Fig. 2. Difjcerenfiafim pnffertrs of CD4’ n~zdCD8& T cells.CD4 ’ nrd CD8 ’ T cells diffrrcntintp
from miue precursors fThpJ, zuhich do not secrete IL-4 or KIN-~, ir~to TIG/TCZ07 T/I~/~CZcd/s.
tion of a whole particle. This effect is exag-
gerated if the pathogen is localized to small Sirnilnr cytokines rrgrrlufe the rliffrentinfiou of CD4 +nmf CD8 r cells.Abbrcz~infio~zs:
[FN-7, infer-
feron y; IL-d, interleukin 4; LT, 1yrnphofoxin; R, T cyfofosic; TGF-fl, t rm$ormirty grox~th@for fi;
foci of infection. Thus, ‘low’ antigen dose can
7?1,T helper; Thp, T helper precursor; Thp’, nnfige~~-sfi~~!~~~~fe[~
proli~wztirg T1zp.
mean a low amount of antigen on all APCs,
or a higher dose on infrequent AJ?Cs. These
two situations may need to be considered separately. cells in humans and mice3@@.CD4’ and CD8+ subsets can be de-
rived under similar conditions: IL-12 and IFN-y encourage the
differentiation of precursors into Thl or Tel cells, whereas IL-4
Mulriplecriteriafbr the ThI-Th2 choice? induces the generation of Th2 or Tc2 cells. Previous difficulty in
The Thl-Th2 decision is crucial for effective immunity; therefore, it identifying Tc2 cells may have been due to the strong bias of
may be advantageous for pathogens to subvert this decision. If any sin- CD8+ T cells towards differentiation into IFN-y producers.
gle criterion (e.g. antigen dose) were solely responsible for the Thl-Th2 Indeed, during initial priming, CDB- T cells require a higher
decision, it is likely that microorganisms, with rapid evolutionary dose of IL-4 than do CD4+ T cells in order to differentiate into
rates, would have evolved ways to interfere. For instance, it is well IL-4-producing cells (S. Sad and T.R. Mosmann, unpublished).
known that viruses have evolved sophisticated mechanisms of im- After commitment of CD8+ T cells either to Tel or to Tc2 pheno-
mune interference based on the ‘theft’ of several immune-related types, neither subset can be converted to the other cytokinc-
genes, including cytokine receptorsJ4js. Thus, it makes sense that the secretion pattern”‘.
decision as to which effector function is required should be based on a Subsets of CD8+ T cells have been identified in humans as well
complex matrix of interlocking factors, since this should make the cru- as in mice during various infections. Tc?-like cells have been
cial choice of effector function more resilient to pathogen interference. isolated from patients with lepromatous !eprosy and from human
immunodeficiency virus (HIV)-infected individuals with a Job’s
like syndrome (high 1gE levels), whereas CD8’ Tel cells have been
Cytokine-secretion subsets of CD8+ 1 cells isolated from patients with tuberculoid leprosy”“,3’. Furthermore,
Tel and Tc2 cells lymphocytic choriomeningitis virus (LCMW-specific CD8 T cells
CD8+ T cells often secrete a Thl-like cytokine pattern. There is secreting IL-5 have been shown to be associated with airway
now increasing evidence for the existence of Th2-like CD8’ T eosinophi1ia4”.

MARCH 1996
 priate B cells to provide cognate help. Foreign antigen peptides pre-
sented by MHC class I would normally indicate that the B cell is in-
fected, and so cytotoxicity would be the appropriate response, instead
of help, However, it remains possible that noncognate, or bystander,
B-cell help by CDS+ Tc2 cells may occur in viva, perhaps due to their
Pe expression of CD4OL (Ref. 41) or secretion of type 2 cytokines, which
aid B cells activated by a CD4’ Th cell (Fig. 3).

Further variants of Tc I cell5

Fig. 3. DO CD&‘+Tcz cd/s kill (red arrows) or help (green arrows) B cells?
The strong cytotoxicity of 7X? cells and the recognition of MHC clnss I-
restricted antigens are both incompatible with cognnte B-cell help. However,
when shmalated by R virus-infected cell (either R B cell or any other cell),
7’c2cells express cytokines and CWOL, zohichmay provide noncognate aug-
mentation of B-cell help provided by Th cells. Abbreviations: Ab, antibody;
Ag, antigen; CD&IL, CD40 ligand; FasL, As &and; n-4, interlettkin 4;
MHC, major histocompatibility complex; R, T cyfotoxic; Th, T helper.
Cytotoxic T lymphocyte (CTL)activity
CDS’ Tc2 cells are as cytotoxic as Tel cells in some38.39,but not
otherr7*41,systems. This may depend on cytokine exposure prior to
the cytotoxicity assav3g.
, The presence or absence of their charac-
Within the Tel subset, further differentiation is possible. Some, but
not all, CD8’ clones secrete IL-2 and, even in the absence of antigen
stimulation, IL-4 rapidly induces Tel clones to lose their ability to
synthesize cytokines, particularly IL-2 (Ref. 43). This appears to be
irreversible, although a similar state (‘anergy’), induced by stimu-
lation in the absence of costimulators, can be reversed by IL-2 treat-
ment44. IL-Ctreated Tel cells retain normal cytotoxic function, but
proliferate after antigen stimulation only in the presence of exogen-
ous T-cell growth factors. Thus, IL-4 abrogates autonomous prolif-
eration of Tel cells, limiting clonal expansion without impairing
short-term effector function. This may be another example of cross-
inhibition between Thl /Tel and Th2/Tc2 responses.
Th I ITh2 patterns: times, locations and contributions
of non-T ceils
Although ThI and Th2 cells are major sources of their respective
cytokines, many other cells within and outside the immune system
also produce these cytokines. Subsets of ~6 T cells, CD4+ and CDB’
cells can all secrete Thl- or ThZ-like cytokme pattenls3g,d5. NK cells
produce IFNy and TNF, and contribute to Thl-like responses25. As
mentioned earlier, IL-4 (and possibly other Th2 cytokines) are
synthesized by mast cells, B cells, basophils and CD3+CD4+NKl.l l

teristic cytokines during cell killing does not appear to affect cyto- cells26*46.
Furthermore, IL-10 is produced by macrophages, keratino-
toxicity (S. Sad et al., unpublished). Tel and Tc2 cells both kill cytes and, as yet unidentified, cells in the placenta34. Thus, several
mainly by a Ca2+/perforin-dependent mechanism, and to a lesser cell types may contribute to an overall Thl or Th2 cytokine pattern
extent via Fas (Ref. 42; S. Sad, L. Krishnan, R.C. Bleackley, D. Kagi, (Table 11, and it has been suggested that these responses should
H. Hengartner and T.R. Mosmann, unpublished). instead be described as type 1 or type 2 (Ref. 36).
Although many interesting Thl or Th2 biases have been found
in infections and autoimmune diseases (Table 21, it is likely that
&elf help careful consideration of anatomical location and the kinetics of the
Although CD4’ Th2 cells show a clear-cut B-cell helper activity and response will provide more definitive data. Many immune re-
anti-CD3-stimulated Tc2 cells can express CD40 ligand (CD4OL), sponses may remain localized; thus, sampling of peripheral blood
CD8’ Tc2 cells do not help B cells in a cognate helper assay39. The lack lymphocytes (PBLs) may provide information on potential re-
of cognate help might be due to their strong cytoiyuc activity: Tel sponses, whereas sampling of lymph node cells is required to rep-
and Tc2 cells can both kill even small, resting B cells (S. Sad, resent ongoing responses. Furthermore, the timing of assay is im-
L. Krishnan, R.C. Bleackley, D. Kagi, H. Hengartner and T.R. portant, as shown by a mouse malaria model in which Thl and Th2
Mosmann, unpublished). However, TCZcells can help B cells in the responses are predominant at different stages of infection4’.
ljresence of platebound anti-CD3 antibody, in which case the cyto-
lybc activity may be masked or focused away from the B cell. Cognate
help by Tc2 cells is also unlikely because of the antigen-processing Functional T-cell s&r&s, surface markers and
pathways. Specific antigen captured by B-cell antibody receptors is memory 1 cells
normaUy processed and presented on MHC class 11and it is unclear Since the discovery of the Thl and Th2 subsets, there has been intense
how CDS’ cells, restricted to MHC class I, could recognize the appro- interest in finding corresponding cell-surface markers that would

MARCH 1996
 IMMUNOLOGY TODAY

allow separation of the subsets from normal cell populations. one of several interactions between immunity and other system, su&
Although initial results suggested that CD45RB was expressed prefer- as wound healing and the nenroendocrine system.
entially on Th2 cells, it is now clear that this cell-surface marker shows
more correlation with naive versus activated/memory cells than with
cytokine phenotype. Thus, naive cells are normally CD45RBhi,but Thl Conclusion
and Th2 cells can both be found in the CD45RBhi or CD4SRB~~ popu- Theexistence of the basic effector Thl and T’h2subsets l.snow well ac-
lations. However, recent data on human T-cell expression of CD30 cepted, and is being used to plan therapeutic and vaccine strategies.
suggests that this might be a useful marker for activated Th2 cells?. However, we do not yet know the full extent of T-cell diversity.
Although Thl and Th2 subsets both express CD30, expression on Th2 Current single-cell assays have proved promising for analyzing cyto-
cells is shonger and more sustained. The potential utility of thii kine expression in normal cells, and three- or four-color fluorescence
marker was shown by the isolation of cells specific for a recently ex- should allow the identification of other phenotypes. A further impor-
posed allergen from the rare CD30’ population. tant question relates to the nature of the initial signal that determines
Another important question is the nature of long-term T-cell the response pattern. In the face of diabolical interference by microbes,
memory. Do memory cells retain the Thl or Th2 commitment of the the success of the immune system in choosing correctly leads us to ex-
active response, or are they uncommitted? Adoptive transfer of trans- pect sophisticated and complex mechanisms for making this decision.
genie T cells into mice lacking endogenous T cells resulted in reten-
tion of a committed Thl or Th2 phenotype for 20 weeks@. However,
under these circumstances, the transgenic T cells are not subjected
to the normal competition of ongoing T-cell responses, which may
be an important regulatory influence that shortens the lifespan of
effector T cell+‘. Activated spleen cells from ‘clean’ mice, with a References
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