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Table of Contents
Introduction ................. Humoral immunity in innate immune system ............ Cellular immunity in the innate immune system ........ Pattern recognition receptors (PRRs) in innate immunity ..... 4 5 6 9

Innate immunity influences adaptive immunity ....... 11 Concluding Remarks ............. 11 References ................... 12

Introduction
mmune system refers to the cells and molecules responsible for immunity, which is a collection of mechanisms within an organism. Healthy individuals protect themselves against diseases by means of an immune system within the body, that functions by identifying and killing a wide variety of pathogens, such as viruses, bacteria, and parasitic worms, as well as tumor cells. The immune system distinguishes foreign material from the organism's normal cells and tissues. Two layers of defense systems are found in organisms, the innate immune system and the adaptive immune system. The innate (also called natural or native) immune system is a front line of defense providing an immediate, but non-specific response found in all plants and animals.[1] A second layer of protection possessed by all vertebrates is provided by the adaptive immune system, which is also called acquired or specific immunity. The adaptive immune system has improved recognition of the pathogen and retains specific responses in the form of an immunological memory that allows the host to mount faster and stronger attacks upon subsequent encounters with pathogens.[2] Both the innate and adaptive immune systems are comprised of both cellular and soluble components (known as humoral immunity) by which they carry out their protective function, but they differ in a number of ways (Table 1). The innate system is thought to constitute an evolutionarily older defense strategy, based on the fact that non-vertebrate organisms use immune systems similar to the vertebrate innate immune system. The major functions of the vertebrate innate immune system include: 1) Recruiting immune cells to sites of infection and inflammation, through the production of specialized chemical mediators, such as cytokines and chemokines. 2) Activation of the complement cascade to identify bacteria, activate cells, and to promote clearance of dead cells or antibody complexes. 3) The identification and removal of foreign substances present in organs, tissues, the blood and lymph, by specialized white blood cells. 4) Activation of the adaptive immune system through a process known as antigen presentation. Both humoral and cellular components of innate immunity are required for these protective functions.

Table 1 : Features of the immune system

Innate immune system

Non-specific response Immediate maximal response Broad receptors recognize PAMPs (Pathogen associated molecular patterns) Receptors are PRRs (Pattern recognition receptors) Relatively stereotypic No memory of prior exposure In all forms of life Components Physical & chemical barriers (Skin, mucosal epithelia & antimicrobial substances, e.g. defensins) Humoral immunity: Complement Cellular immunity: Phagocytes, NK cells

Adaptive immune system

Antigen-specific response Lag time between exposure and maximal response Narrow specific receptors recognize a particular epitope Receptors are B-cell (BCR) and T-cell (TCR) receptors Highly specialized Memory of prior exposure Vertebrates only

Cutaneous & mucosal immune system secreted antibodies Humoral immunity: antibodies Cellular immunity: Lymphocytes

Humoral immunity in innate immune system

nce infectious agents have breached the anatomical barriers and penetrated tissues, acute inflammation is stimulated and is typically manifested by symptoms of redness, swelling, heat, pain, and possible dysfunction of the organs or tissues involved. Humoral factors play an important role in inflammatory edema and the recruitment of phagocytic cells. These humoral factors (see: Table 2) are found in serum or they are formed at the site of infection.

Table 2 : Humoral Factors in the innate immune system

Humoral components
Complement, Mannose-binding lectin Coagulation Platelets Lactoferrin and Transferrin Lysozyme Fibronectin Interferons Interleukin-1 TNF Defensins

Effector function
Opsonization, enhanced phagocytosis, inflammation Chemotactic agents for phagocytic cells, inflammation Antimicrobial substance, e.g. beta-lysin Iron binding, limiting bacterial growth Peptidoglycan hydrolysis, breaking down the cell wall of bacteria Opsonization and phagocytosis Antiviral proteins Fever, Opsonization antiviral, phagocyte activation Antibacterial, antiviral proteins

The complement system is the major humoral component of innate defense mechanism mention.
CLASSICAL PATHWAY Antigen:antibody complexes (pathogen surfaces) C1q, C1r, C1s C4 C2 MB-LECTIN PATHWAY Mannose-binding lectin binds mannose on pathogen surfaces MBL, MASP-1, MASP-2 C4 C2 ALTERNATIVE PATHWAY Pathogen surfaces C3 B D

[3, 4]

and deserves further

C3 convertase Terminal complement components C5b C6 C7 C8 C9 Membrane-attack complex, lysis of certain pathogens and cells

C3a, C5a

C3b

Peptide mediators of inflammation, phagocyte recruitment

Binds to complement receptors on phagocytes Opsonization of pathogens Removal of immune complexes

Figure 1a Overview of the main components and effector actions of complement

The complement system is a that attacks the surfaces of foreign cells. The term "complement" was introduced by Paul Ehrlich in the late 1890s, who described it is something in the blood which "complements" the cells of the immune system in the killing of pathogens. The complement system is made up of over 25 different serum small proteins and protein fragments, including serum proteins, serosal proteins, and cell membrane receptors. These proteins are synthesized mainly in the liver, and they account for about 5% of the globulin fraction of blood serum. Complement proteins circulate in the blood in an inactive form. Elements of the complement cascade can be found in many species evolutionarily older than mammals including plants, birds, fish , some species of invertebrates[5], as well as in humans. The complement system is not adaptable and does not change over the course of an individual's lifetime; as such it belongs to the innate immune system. However, it can be recruited and brought into action by the adaptive immune system . After complement proteins initially bind to microbes, they activate their protease activity, which in turn activates other complement proteases, and so on. This produces a catalytic cascade that amplifies the initial signal by controlled positive feedback.[6, 7] The cascade results in the production of peptides that attract immune cells, increase vascular permeability, and opsonize (coat) the surface of a pathogen, marking it for destruction. This deposition of complement can also kill cells directly by disrupting the plasma membrane barrier.[3]

Functional protein classes in the complement system

Binding to antigen:antibody complexes and pathogen surfaces Binding to mannose on bacteria C1q MBL C1r C1s C2b Bb D MASP-1 MASP-2 C4b C3b C5a C3a C4a C5b C6 C7 C8 C9 CR1 CR2 CR3 CR4 C1qR C1INH C4bp CR1 MCP DAF H I P Cd59

Activating enzymes

Membrane-binding proteins and opsonins Peptide mediators of inflammation

Membrane-attack proteins

Complement receptors

Complement-regulatory proteins

Figure 1b Functional protein classes in the Three biochemical pathways activate the complement complement system. system: the classical complement pathway, which initiates complement activation via binding to specific antibodies; the alternative complement pathway, which activates via direct binding to pathogen surfaces; and the mannose-binding lectin pathway, which is homologous to the classical complement pathway, but involves a protein that binds to mannose residues and other sugars on multiple pathogens.[5] Figure 1a & b diagrammatically show the major players in all three complement activation pathways of vertebrates [the diagrams are adopted from Janeway CA, Jr. et al (2005). Immunobiology., 6th ed., Garland Science].

Cellular immunity in the innate immune system

he cellular component of the innate immune system consists of a number of white blood cells (WBC), also known as leukocytes, which includes: Natural killer cells, mast cells, eosinophils, basophils, and phagocytic cells, including macrophages, neutrophils and dendritic cells. They are all derived from CD34+ pluripotent hematopoietic stem cells present in the bone marrow.[8] See Table 3 for characteristics of cellular components involved in the innate immune system.

Since leukocytes are able to move freely, part of the inflammatory response is their recruitment to sites of infection. These cells are the main line of defense in the non-specific immune system by interacting, identifying, capturing cellular debris, foreign particles or invading microorganisms, and eliminating the pathogens that might cause infection.[5] Phagocytic cells in the innate immune system utilize a number of processes to carry out their function, including chemotaxis to sites of infection, production of chemicals, enzymes, chemokines and cytokines that initiate inflammation and other immune responses, as well as induce apoptosis and engulf infected cells. [5, 8, 11] In addition, macrophages and dendritic cells also serve as professional antigen-presenting cells (APCs) that display a fragment of foreign antigen complexed with MHC II molecule on their surface. Cells from adaptive immune system, such as T cells, recognize and interact with the antigen-class II MHC molecule complex on the membrane of the antigen presenting cell. An additional co-stimulatory signal such as B7 is then produced by the antigen presenting cell, leading to activation of the T cell which triggers adaptive immunity. [8, 11] Neutrophils, eosinophils and basophils are classified together as granulocytes. Neutrophils are the most common leukocyte type in the blood and are the major responders to inflammation and infection. Eosinophils and basophils along with mast cells are often associated with allergic reactions, but normally carry out protective response in killing pathogens such as parasites. Natural killer (NK) cells also known as large granular lymphocytes (LGL) because they resemble lymphocytes in their morphology, except that they are slightly larger and have numerous granules in their cytoplasm contain special proteins such as perforin and proteases known as granzymes.[5] They can be identified by the presence of CD56 and CD16 and a lack of CD3 cell surface markers. NK cells can nonspecifically kill viral infected cells and tumor cells by releasing perforin that forms pores in the cell membrane of the target cell through which the granzymes and associated molecules can enter, inducing apoptosis of infected cells and preventing the spread of virus. [22] The killing becomes more efficient upon exposure to IL-2 and IFN, wherein NK cells become lymphokine-activated killer (LAK) cells, which are capable of killing malignant cells. Continued exposure to IL-2 and IFN enables the LAK cells to kill transformed as well as malignant cells. It should be noted that anti-viral infection reactions and tumor surveillance of NK cells are not inflammatory responses. For a long time, researchers questioned how NK cells can distinguish a normal cell from a virus-infected or malignant cell. Now we know that NK cells have two kinds of receptors on their surface a killer activating receptor (KAR) and a killer inhibiting receptor (KIR). When the KAR encounters its ligand, a killer activating ligand (KAL) on the target cell, the NK are capable of killing the target. However, if the KIR also binds to its ligand then killing is inhibited even if KAR binds to KAL. The ligands for KIR are MHC class I molecules. So, as long as a target cell expresses class I MHC molecules, it will not be killed by NK or LAK cells even if it expresses KAL. Normal cells constitutively express MHC class I molecules on their surface, however, virus infected and malignant cells down regulate expression of class I MHC. Thus, NK and LAK cells selectively kill virus-infected and malignant cells while sparing normal cells. [29-32] Cells capable of killing foreign and altered self target cells in a non-specific manner are also granted a name, Killer (K) cells. NK cells, activated macrophages and eosinophils belong to this group of cells. Killer cells that carry Fc receptors like NK and macrophages for IgG antibodies and Fc receptor like eosinophils for IgE antibodies can mediate antibodydependent cellular cytotoxicity (ADCC). In ADCC, bound antibodies direct the K cells to the target cells, initiating cell killing. These cells play an important role in the innate immune system. [22] g d T cells are here counted in innate immune system because they exhibit characteristics that place them at the border between innate and adaptive immunity. One on hand, T cells may be considered a component of adaptive immunity in that they rearrange TCR genes to produce junctional diversity and develop a memory phenotype. However, the various subsets may also be considered part of the innate immune system where a restricted TCR or NK receptors may be used as a pattern recognition receptor. [23-28]

Table 3 : Cellular components involved in the innate immunity system

Cell
Mast cells

Function
against pathogens, wound healing, allergy and anaphylaxis, recruits neutrophils and macrophages, dilates blood vessels

Product
histamine heparin chemokines, or chemotactic cytokines

Location
connective tissue and mucous membranes

Reported Markers

Reference
[9-10]

Activation
IgE:allergen attachment

Receptors
FcRI SCF receptor (ckit, Cd117)

CD9, CD33, CD43, CD45, CD54, CD117 (c-kit, SCF receptor), HLA-class I, IgE Fcg Rs, CD11c,CD14, CD16/CD32, CD68, CD82, CD163 HLADM, MHC class

Macrophages

Large phagocytic cells, chemotaxis, phagocytosis destroy bacteria through respiratory burst, inflammation, apoptosis, antigen presentation

reactive oxygen species, Cathepsins, lysozyme, PLA2, iNOS, MMPs (1, 2, 7, 9, 12), chemokines/cytoki nes (IL-8, Rantes, MIP1a, MIP1b, MIP-2, IP-10, IFN, IL-1a, IL-1b, TNF, IL-6, IL-12, TGFb, PDGF, FGFb, VEGF, IGF), Fas Ligand oxidizing agents including hydrogen peroxide, free oxygen radicals and hypochlorite, Defensins and the serine proteases neutrophil elastase and cathepsin G

Moving to the areas between cells in pursuit of invading pathogens.

[5, 18, 19]

LPS, LBP IFNg , MSP, IL4, IL-13,

MHC class II, Fcg receptors (I-III), B7-1, B7-2, Scavenger receptors, Complement receptors, MSPR,TLR4, IFNg Rs, CXCRs, IL-4Ra, IL-13Ra1

Neutrophils, most abundant type of phagocyte

phagocytic cells, chemotaxis, phagocytosis, destroy bacteria through respiratory burst

the first cells to arrive at the site of an infection.

CD11b, CD15, CD66, CD66b, Ly-6G (for mouse)

[11, 12, 17]

IL-8, IFNg C5a,

b 2-integrins, Fcg receptors, lselectin, complement receptor 1 (CR-1), C5a receptor, decay-accelerating factor (DAF), intercellular adhesion molecule-1 (ICAM-1) and ICAM-3

Dendritic cells [Myeloid dendritic cells (mDC), Plasmacytoid dendritic cells (pDC)]

phagocytic cells, phagocytosis, antigen presentation (TH cell activators) serving as a link between the innate and adaptive immune systems

Cytokines (IL-12 by mDC, IFN-a by pDC)

Skin (Langerhans cells), mucosa

CD1a, CD11c, CD40, TLR 2 & TLR 4 (mDC), TLR 7 & TLR 9 (pDC) The markers BDCA-2, BDCA-3, and BDCA-4 for discriminate among the types

[8, 13, 16]

MHC class II, B7s, TLRs, LTb Rs TLRs, BDCA-2, BDCA-3, BDCA-4, CD40LTb Rs

Basophils Eosinophils

against parasites, allergic reactions (such as asthma)

histamine toxic proteins and free radicals

Infectin site Infectin site

Ly-6G (for mouse)

[5, 21] [11, 20]

FcR TLR FcR

killing bacteria and parasites, allergic reactions attack and destroy tumor cells, and virally infected cells by inducing apoptosis bridge between innate and adaptive responses

Cd15, Ly-6G (for mouse), CD67

Human: CD16, CD56, CD94/NKG2A/B Mouse: Ly49, NK1.1/NK1.2

Natural killer cells, or large granular lymphocytes

perforin and granzymes, cytokines (IFN, IL-4)

Infectin site

[5, 22, 2932]

IFNa /b , IL-2, IFN, IL-12, Cd1d

FcR, KAR, KIR, CD94/ NKG2, Ly49, LIR

g d T cells

abundance in the gut mucosa

g d TCR, CD94, Vg 9/Vd 2

[23-28]

PRR: Vg 9/V2

g d TCR, CD94, Vg 9/V2

Pattern recognition receptors (PRRs) in innate immunity

hagocytic cells have a variety of receptors on their cell membranes that recognize distinct molecular patterns on infectious agents. There are three groups of PRRs: 1) Secreted PRRs, 2) Phagocytic receptors and 3) Toll-like receptors (TLRs). Secreted PRRs are molecules that circulate in blood and lymph system such as complement. Phagocytic cells have a receptor for the 3rd component of complement, C3b. The binding of C3b-coated bacteria to this receptor results in enhanced phagocytosis and stimulation of respiratory burst. Phagocytic cells also have surface receptors that bind the pathogen for engulfment, such as Fc receptors and Scavenger receptors. Toll-like receptors are cell surface or intracellular transmembrane receptors that bind to pathogens or other pathogen-associated material and initiate signaling leading to the release of cytokines. These receptors were so-called Toll-like receptors due to their homology to receptors first discovered and named in Drosophila. [33] Macrophages and dendritic cells have a set of Tolllike receptors that recognize broad types of PAMPs on infectious agents. Binding of infectious agents to Toll-like receptors recruits intracellular adapter molecules that propagate a signal. Four adapter molecules are known to be involved in TLR signaling. These proteins are known as MyD88, Tirap (also called Mal), Trif, and Tram.[7, 8, 37-39] The adapters activate other molecules within the cell, including certain protein kinases (IRAK1, IRAK4, TBK1, and IKKi) that amplify the signal. The TLRs will then identify the nature of the pathogen and turn on an effector response. These signaling cascades direct the expression of various cytokine genes and the release of inflammatory cytokines (IL-1, TNF-alpha and IL-6) by phagocytes, ultimately leading to the induction or suppression of genes that orchestrate the inflammatory response. In all, thousands of genes are activated by TLR signaling, and collectively, the TLRs constitute one of the most powerful and important gateways for gene modulation. Figure 2 diagrams the signaling pathway of TLRs.

Figure 2 : Signaling pathway of toll-like receptors

In mammals, there are about 13 TLRs reported so far[34-36], each of which, specializes in a subset of PAMPs. See Table 4 for the summary of known Mammalian Toll-like Receptors.
Table 4 : Summary of Mammalian Toll-like Receptors
Receptor
TLR 1

Ligand(s)
triacyl lipoproteins lipoproteins; gram positive peptidoglycan; lipoteichoic acids; fungi; viral glycoproteins

Target Pathogen(s)

Adapter(s)
MyD88/MAL

Location
cell surface cell surface

TLR 2

Gram-positive bacteria (e.g. Streptococci, Staphylococci), Viruses

MyD88/MAL

TLR 3

double-stranded RNA (as found in certain viruses), poly I:C

Viruses

TRIF

cell compartment

TLR 4

lipopolysaccharide (endotoxin); viral glycoproteins

Gram-negative bacteria (e.g. Salmonella, E. coli O157:H7)

MyD88/MAL/TRIF/TRAM

cell surface

TLR 5

flagellin

Mobile bacteria (e.g. Listeria)

MyD88

cell surface

TLR 6 (Forms a heterodimer with TLR-2)

diacyl lipoproteins, certain lipids

MyD88/MAL

cell surface

TLR 7

small synthetic compounds; singlestranded RNA

ssRNA viruses (e.g. Influenza, Measles, Mumps)

MyD88

cell compartment

TLR 8

small synthetic compounds; singlestranded RNA

ssRNA viruses (e.g. Influenza, Measles, Mumps)

MyD88

cell compartment

TLR 9

unmethylated CpG DNA

CpG DNA pathogen

MyD88

cell compartment

TLR 10

unknown

unknown

cell surface

TLR 11 (not in human)

Profilin

Protozoans (Apicomplexa)

MyD88

cell surface

TLR 12 (not in human)

unknown

unknown

unknown

TLR 13 (not in human)

unknown

unknown

unknown

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Innate immunity influences adaptive immunity

he innate immune system does not act separately from the adaptive immune system. In fact, the innate immunity can trigger adaptive immunity. This can occur in several ways. 1) Macrophages and dendritic cells are phagocytes that are also responsible for "presenting" antigens to T cells to initiate both cell-mediated and antibody-mediated adaptive immune responses. 2) The interaction of PAMPs and TLRs on dendritic cells causes them to secrete cytokines, including interleukin 6 (IL-6), which interfere with the ability of regulatory T cells to suppress the responses of effector T cells to antigen. 3) Pathogens coated with fragments of the complement protein C3 are not only opsonized for phagocytosis but also bind more strongly to B cells that have bound the pathogen through their BCR. This synergistic effect enables antibody production to occur at doses of antigen far lower than would otherwise be needed. 4) B cells are also antigen-presenting cells that function similarly to macrophages and dendritic cells. They bind antigen with their BCRs and engulf it into lysosomes, where the digested fragments are incorporated into class II MHC molecules, which are then presented to T cells. In addition, B cells also have TLRs. When a PAMP such as LPS binds the TLR, it enhances the response of the B cell to the antigen. All in all, adaptive immunity is not possible without the assistance of the mechanisms of innate immunity.

Concluding remarks
he innate immune system, as a first line of defense against pathogens, involves a diverse collection of molecules and mechanisms that include chemicals, enzymes, cytokines, chemokines, and effector cells (see tables 1-4). Overall, many different cellular processes are involved, including cell signaling via receptors, phagocytosis, targeted cell killing, and apoptosis. BioLegend provides a variety of reagents, antibodies, and tools for study of the innate immune system including leukocyte phenotyping, cytokine and chemokine measurements, receptor detection, measurement of apoptosis and cell signaling molecules.

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21. Komiya A (2006) Expression and Function of Toll-Like Receptors in Human Basophils International Archives of Allergy and Immunology 2006;140 (Suppl. 1):23-27. 22. Bots M, Medema JP (2006). "Granzymes at a glance". J. Cell. Sci. 119 (Pt 24): 5011-4. 23. Holtmeier W, Kabelitz D. T cells link innate and adaptive immune responses. Chem Immunol Allergy. 2005;86:151-183. 24. Born WK, Reardon CL, O'Brien RL. The function of T cells in innate immunity. Curr Opin Immunol. 2006;18:3138. 25. Morita CT, Mariuzza RA, Brenner MB. Antigen recognition by human T cells: pattern recognition by the adaptive immune system. Springer Semin Immunopathol. 2000;22:191-217. 26. Eberl M, Hintz M, Reichenberg A, Kollas AK, Wiesner J, Jomaa H. Microbial isoprenoid biosynthesis and human T cell activation. FEBS Lett. 2003;544:4-10. 27. Girardi M. Immunosurveillance and immunoregulation by T cells. J Invest Dermatol. 2006 Jan;126:25-31. 28. Thedrez A, Sabourin C, Gertner J, Devilder MC, Allain-Maillet S, Fournie JJ, Scotet E, Bonneville M. Self/non-self discrimination by human T cells: simple solutions for a complex issue? Immunol Rev. 2007;215:123-135. 29. Lieto LD (2006) The human CD94 gene encodes multiple, expressible transcripts including a new partner of NKG2A/B Genes and Immunity 7, 3643. 30. Makrigiannis AP (2001) Class I MHC-binding characteristics of the 129/J Ly49 repertoire. J Immunol. 2001 Apr 15;166(8):5034-43. 31. Butsch Kovacic et al. (2005) Variation of the Killer Cell Immunoglobulin-Like Receptors and HLA-C Genes in Nasopharyngeal Carcinoma. Cancer Epidemiol Biomarkers Prev 14 (11): 2673. 32. Gonen-Gross et al. (2005) 175 (8): 4866. The CD85J/Leukocyte Inhibitory Receptor-1 Distinguishes between Conformed and _2-Microglobulin-Free HLA-G Molecules. The Journal of Immunology 175: 48664874. 33. Lemaitre,B., Nicolas,E., Michaut,L., Reichhart,J.M., and Hoffmann,J.A. 1996. The dorsoventral regulatory gene cassette spatzle/Toll/cactus controls the potent antifungal response in Drosophila adults. Cell 86:973-983. 34. Du,X., Poltorak,A., Wei,Y., and Beutler,B. 2000. Three novel mammalian toll-like receptors: gene structure, expression, and evolution. Eur. Cytokine Netw. 11:362-371. 35. Chuang,T.H., and Ulevitch,R.J. 2000. Cloning and characterization of a sub-family of human toll-like receptors: hTLR7, hTLR8 and hTLR9. Eur. Cytokine Netw. 11:372-378. 36. Tabeta,K., Georgel,P., Janssen,E., Du,X., Hoebe,K., Crozat,K., Mudd,S., Shamel,L., Sovath,S., Goode,J. et al 2004. Toll-like receptors 9 and 3 as essential components of innate immune defense against mouse cytomegalovirus infection. Proc. Natl Acad. Sci. U. S. A 101:3516-3521. 37. Yamamoto,M., Sato,S., Hemmi,H., Hoshino,K., Kaisho,T., Sanjo,H., Takeuchi,O., Sugiyama,M., Okabe,M., Takeda,K. et al 2003. Role of adapter TRIF in the MyD88-independent Toll-like receptor signaling pathway. Science 301:640-643. 38. Yamamoto,M., Sato,S., Hemmi,H., Uematsu,S., Hoshino,K., Kaisho,T., Takeuchi,O., Takeda,K., and Akira,S. 2003. TRAM is specifically involved in the Toll-like receptor 4-mediated MyD88-independent signaling pathway. Nat. Immunol. 4:1144-1150. 39. Yamamoto,M., Sato,S., Hemmi,H., Sanjo,H., Uematsu,S., Kaisho,T., Hoshino,K., Takeuchi,O., Kobayashi,M., Fujita,T. et al 2002. Essential role for TIRAP in activation of the signalling cascade shared by TLR2 and TLR4. Nature 420:324-329.

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