Measurement of mouse and human interleukin 10.
(Curr Protoc Immunol. 2001 May;Chapter 6:Unit 6.14.)
University of Alberta, Edmonton, Alberta, Canada.
Interleukin 10 (IL-10) is a multifunctional cytokine produced by T cells and
other cell types. It regulates activities of a variety of the cells of the immune
system and may play a crucial role in cross-regulation of different types of T
cells. Two methods of quantitating IL-10 are provided in this unit. The first is
an enzyme-linked immunosorbent assay (ELISA) using a pair of monoclonal
antibodies specific for mouse IL-10. This method has the advantages of high
specificity and insensitivity to other cytokines present in test samples. The
second method is a bioassay which has the advantages that both human and mouse
IL-10 are detected and that functional protein is measured. Both methods have
Measurement of human interleukin 11.
(Curr Protoc Immunol. 2001 May;Chapter 6:Unit 6.15.)
Bennett F, Gianotti J, Celniker A, Turner KJ, Clark SC.
Genetics Institute, Inc., Cambridge, Massachusetts, USA.
This unit describes an ELISA and a cell proliferation assay that can be used,
respectively, to measure the protein level or biologic activity of human and
murine interleukin 11 (IL-11). The bioassay is based on the ability of IL-11 to
support growth of the B9-11 cell line, a subline of B9 that has traditionally
been used to measure levels of IL-6. B9-11 is substantially more responsive to
IL-11 than the T10 line used in older protocols. This new bioassay therefore
provides improved sensitivity, with a detection limit of 20 pg/ml. An alternate
procedure is provided that employs neutralizing antibodies in the cell
proliferation bioassay to use to ensure that the activity of the desired molecule
(IL-11) is being measured in samples containing multiple cytokines. A describes
maintenance of B9-11 cells.
Measurement of interleukin 15.
(Curr Protoc Immunol. 2001 May;Chapter 6:Unit 6.22.)
Immunex Corporation, Seattle, Washington, USA.
Interleukin 15 (IL-15) is a small helical cytokine that was first characterized
by its ability to stimulate proliferation of the murine T cell line CTLL-2, but
is now known to have activity on a variety of cell types. This unit describes two
protocols that can be used to measure IL-15 in culture supernatants and serum
samples. The first method uses an enzyme-linked immunosorbent assay (ELISA) that
is specific for human IL-15, although it can also detect simian IL-15. The second
method relies on the ability of IL-15 to support proliferation of CTLL-2 cells.
The CTLL-2 assay is slightly more sensitive than the ELISA, but it is not
specific for IL-15, because CTLL-2 cells also respond to IL-2 and IL-4. The
CTLL-2 assay can be used to measure human, simian, or murine IL-15.
Measurement of interleukin 16.
(Curr Protoc Immunol. 2001 May;Chapter 6:Unit 6.23.)
Center DM, Cruikshank WW, Parada NA, Ryan T, Theodore AC, Viglianti G, Lim KG,
Boston University Medical Center, Boston, Massachusetts, USA.
Interleukin 16 (IL-16) is a chemoattractant immunomodulatory cytokine that
initiates its cellular responses through interaction with membrane-expressed CD4.
The protein may be detected by a number of methods; the choice of protocol will
depend on the ultimate object of a particular experiment. The first method
presented is the use of ELISA to measure IL-16 in cell culture supernatants or
biological fluids. For some applications, such as identification of IL-16 in an
unknown fluid or medium or direct assessment of its bioactivity, functional
assays of IL-16-induced responses may be more appropriate. The chemotactic
effects of IL-16 on CD4+ T cells and its specific inhibition may be measured
using anti-IL-16 antibodies; the same approach may also be applied to monocytes
or eosinophils. Another effect of IL-16 is the induction of CD25, which can be
assayed using immunological staining. Finally, cell cycle progression in target
cells can be measured by the incorporation of radiolabeled thymidine and
confirmed by inhibition with neutralizing antibody.
Measurement of human and mouse interleukin 18.
(Curr Protoc Immunol. 2001 Nov;Chapter 6:Unit 6.26.)
Yoshimoto T, Tsutsui H, Okamura H, Nakanishi K.
Hyogo College of Medicine, Hyogo, Japan.
IL-18, originally designated as interferon-gamma (IFN-gamma)-inducing factor
(IGIF), is a pleiotropic cytokine secreted by activated macrophages and Kupffer
cells. The major activity associated with this cytokine is induction of IFN-gamma
production from T cells, B cells, and NK cells, especially in collaboration with
IL-12. IL-18 is synthesized without a signal peptide and must be enzymatically
cleaved to become active. Therefore, it is important to determine whether the
produced IL-18 is an active or precursor form. This unit describes functional
assays for measurement of bioactive human and mouse IL-18 and ELISAs for
measurement of murine and human IL-18 proteins. The functional assays are based
on the induction of IFN-gamma production by IL-18. The ELISA measures the
concentration of human or mouse IL-18. Using a combination of monoclonal
antibodies against human or mouse IL-18, the proform and/or mature form of IL-18
can be detected by ELISA.
Measurement of mouse and human interleukin 5.
(Curr Protoc Immunol. 2001 May;Chapter 6:Unit 6.5.)
National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.
Two fundamentally different types of assays are available for quantitating
interleukin 5 in biological samples. One type, a bioassay, is based on the
ability of IL-5 to enhance B cell proliferation and immunoglobulin secretion or
eosinophil proliferation and differentiation. The other type of assay, an ELISA,
uses antibodies against IL-5 to capture and quantitate IL-5 in samples.
Advantages and disadvantages of each assay are discussed. The bioassay described
in this unit, utilizing BCL(1) cells as the indicator line, has been designed
primarily to assay murine IL-5; however, it can also be used to measure human
IL-5. The IL-5 ELISA, while sensitive and specific, provides no information about
Measurement of interleukin 6.
(Curr Protoc Immunol. 2001 May;Chapter 6:Unit 6.6.
Nordan RP, Richards CD, Gauldie J.
National Cancer Institute, Bethesda, Maryland, USA.
Interleukin 6 (IL-6) is a pluripotent cytokine with multiple effects on many
different cell types. It is produced by a variety of cells in response to
immunological and other stimuli. This unit describes a simple and sensitive assay
for human, rat, rabbit, pig, and mouse IL-6, based on IL-6-dependent
proliferation of a murine B cell hybridoma cell line, B9. Support protocols
discuss maintenance of B9 cells, preparation of IL-6 standards, and production of
IL-6-containing supernatant. In addition, IL-6 ELISA kits for the measurement of
human or mouse IL-6 are available from a number of companies. These products are
reliable, highly sensitive, and specific, and thus should be considered as an
excellent (although more expensive) alternative, keeping in mind that bioactivity
is not assessed with this approach.
Measurement of interleukin-13.
(Curr Protoc Immunol. 2002 Feb;Chapter 6:Unit 6.18.)
Lauder A, McKenzie AN.
MRC Laboratory of Molecular Biology, Cambridge, United Kingdom.
This unit describes two protocols that can be used to quantitate interleukin-13
(IL-13). The enzyme-linked immunosorbent assay (ELISA) has the advantage of being
highly specific for human IL-13 and does not recognize other cytokines present in
the sample. A bioassay is also presented based on the ability of IL-13 to
stimulate the growth of the B9 plasmacytoma cell line. The bioassay method can be
used to detect both mouse and human IL-13. B9 cells are dependent on IL-6 for
growth and will also respond to IL-4. Thus, although B9 cells can readily be used
to quantify IL-13 in the absence of other cytokines, neutralizing antibodies must
be incorporated into the bioassay when other cytokines are present in the sample.
Measurement of interleukin-17.
(Curr Protoc Immunol. 2007 Nov;Chapter 6:Unit 6.25.)
Pappu BP, Dong C.
M.D. Anderson Cancer Center, Houston, Texas, USA.
Upon antigenic stimulation, naive CD4+ T cells undergo proliferation and
differentiate into cytokine-producing T helper (T(H)) effector cells. T(H)1 cells
secrete effector cytokine IFN-gamma and regulate cell-mediated immunity, whereas
T(H)2 cells produce IL-4, IL-5, and IL-13 cytokines, and mediate immunity against
extracellular pathogens and allergic reactions. Recent studies have identified a
novel T(H) subset, called T(H)17, TH(IL-17), or inflammatory T(H) (THi) cells,
characterized by the production of a proinflammatory cytokine, IL-17, and
regulating inflammatory responses. In this unit, we describe the protocols for
the differentiation of mouse IL-17-expressing T cells in vitro, detection of
IL-17-expressing T cells by intracellular cytokine staining, and measurement of
IL-17 secretion in culture supernatants by ELISA. Generation of IL-17-expressing
T cells in vitro under defined culture conditions allows investigation of their
differentiation regulation. Detection of IL-17 in cell culture and tissue samples
helps in monitoring inflammatory diseases and determining efficacy of therapeutic
Measurement of interleukin-21.
(Curr Protoc Immunol. 2007 Aug;Chapter 6:Unit 6.30.
Zeng R, Spolski R, Leonard WJ.
National Institutes of Health, Bethesda, Maryland, USA.
This unit describes three procedures for measurement of interleukin-21 (IL-21).
The first employs the use of an antibody sandwich ELISA. An alternative procedure
measures proliferative responses of T cells to a combination of IL-21 and IL-15
using CFSE. Finally, a method to assess IL-21-induced tyrosine phosphorylation of
Stat3 in splenic CD8(+) T cells using a flow cytometry-based analysis is
The International Cytokine Society ()
Official web site of cytokine research community ICS.
Cytokines and interleukines
Cytokines are a group of small intercellular signaling molecules (8-30 Kd) synthesized by a variety of tissue and cells. Once bind to cytokine receptors, cytokines exert their functions through autocrine, paracrine or endocrine pathways.
Figure. Cytokine receptors belong to families of receptor proteins, structure (Immunobiology)
Part IV. The Adaptive Immune Response 8. T Cell-Mediated Immunity General properties of armed effector T cells.
Table. Common human cytokines and their receptors (Eurekah Bioscience Collection)
RNA Cytokines, Chemokines and their Receptors Cytokines, their Receptors and their Genes
Cytokines, their Receptors and their Genes (Eurekah Bioscience Collection)
RNA Cytokines, Chemokines and their Receptors