export_immunology test 1 Flashcards

1
Q

Variolation

A

Inoculation of a small amount of pus or scaberous material from an infected individual

Goal= induce a milder form of the disease

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2
Q

Herd immunity

A

When the majority of a population is immune it significantly reduces pathogen reservoir and lowers probability of an uninfected person contracting the disease

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3
Q

Innate immunity

A

In place prior to infection

Rapid

Early defense

Responds same way to repeated exposures

Recognizes general structures of microbes

Limited diversity

Barriers: skin, mucosal epithelia, antimicrobial chemicals

Blood protein: complement

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4
Q

Adaptive immunity

A

Provided by T and B lymphocytes

Response improves w/ subsequent exposure

Highly specific recognition system

Results in immunological memory

Large diversity

Barrier: lymphocytes in epithelia, Ab secreted at epithelial surfaces

Blood proteins: Ab

Cells: lymphocytes

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5
Q

Phagocytes, NK cells

A

Examples of cells of innate immunity

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6
Q

B lymphocytes, t lymphocytes, antibodies, effector t cells

A

Examples of cells of adaptive immunity

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7
Q

Antigen

A

Any foreign substance that induces a specific immune response or is a target of such a response

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8
Q

Humoral immunity

A

Mediated by B cells and results in antibody production and is most effective against extracellular microbes and their toxins

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9
Q

Cell mediated immunity

A

Mediated by T cells and results in activation of phagocytes or killer cells and is most effective against intracellular microbes

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10
Q

Humoral immunity

A

Extracellular microbes

B lymphocytes respond

Effector mechanism: secreted Ab

Transferred by serum

Functions: Block infections and eliminate extracellular microbes

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11
Q

Cell-mediated immunity

A

Microbes: Phagocytosed microbes in macrophage

Responding lymphocyte: Helper T lymphocyte

Effector mechanism: T lymphocytes

Function: Activate macrophages to kill phagocytosed microbes

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12
Q

Cell-mediated immunity

A

Microbe: intracellular microbes replicating w/ in infected cell

Responding lymphocyte: Cytotoxic T lymphocyte

Effector mechanism: T lymphocytes

Function: Kill infected cells and eliminate reservoirs of infection

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13
Q

Active immunity

A

Induced by exposure to a foreign antigen

Has memory

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14
Q

Passive immunity

A

Induced by the transfer of serum or lymphocytes from a specifically immunized individualt to a naive individual eg maternal Ab or antiserum

No memory

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15
Q

Specificity, diversity, memory, clonal expansion, specialization, contraction and homeostasis, nonreactivity to self

A

Cardinal features of adaptive immune response (7)

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16
Q

lymphocyte repertoire

A

Total number of antigenic specificities of the lymphocyte of an individual is called?

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17
Q

Immunologically naive

A

Individuals and lymphocytes that have not encountered a particular antigen are?

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18
Q

B lymphocytes

A

Only cells capable of producing Ab

Recognize extracellular antigens

Differentiate into plasma cells

Mediate humoral immunity

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19
Q

T lymphocytes

A

Only recognize protein antigens presented to them by antigen presenting cells in the context of major histocompatability complex molecules

Provide help to other cells in the form of cytokines, or become killer cells which kill infected cells

Mediate cell mediated immunity

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20
Q

Regulatory T cells

A

Mainly function to inhibit or limit immune responses

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21
Q

Natural Killer cells

A

Involved in innate immunity to viruses

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22
Q

Antigen presenting cells

A

Dendritic cells, macrophages, B cells

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23
Q

B lymphocyte

A

Responds to microbes

Effector function: Neutralization of microbe, phagocytosis, complement activation

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24
Q

Helper T lymphocyte

A

Effector functions: activation of macrophages, inflammation, activation of T and B lymphocytes

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25
Q

Cytotoxic T lymphocytes

A

Recognizes infected cell expressing microbial antigen

Kills infected cell

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26
Q

Regulatory T lymphocyte

A

Suppresses immune response

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27
Q

Natural killer cell

A

Kills infected cell

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28
Q

Cytokines

A

Large and heterogenous group of secreted proteins produced by many cell types that mediate and regulate many aspects of innate and adaptive immunity

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29
Q

Cytokines

A

May be inactive/active

Can be pleiotrophic,redundant, antagonize or synergize

Autocrine, paracrine, or endocrine

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30
Q

Innate immune response

A

Barriers: Skin and mucosa - continuous epithelia

Inflammation: Recruitment and activation of cells and plasma proteins from the blood to kill microbes

Antiviral defense: Cytokine- mediated response in which cells aquire resistance to viral infection, and killing of virus-infected cells by NK cells

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31
Q

Adaptive immune response

A

Ab- bind to extracellular microbes, block their ability to infect host cells, promote their ingestion and destruction by phagocytes

Phagocytes: Ingest microbes and kill them, helper T cells enhance microbicidal abilities of phagocytes

Cytotoxic T cells: Kill cells infected by microbes that are inaccessible to Ab and phagocytic destruction

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32
Q

Dendritic cells

A

Located in epithelia

Capture microbes and antigens

Travel thru lymphatic system to draining lymph nodes

Present antigen to naive T cells

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33
Q

naive B cells

A

WHole antigens or microbes in lymph are captured by ? in the lymph nodes

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34
Q

Helper T cells

A

Help other cells by secreting cytokines

Some remain in secondary lymphoid organs and provide cytokines to aid in activation of B cells while others exit organs and travel to site of infection and help to activate phagocytic cells

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35
Q

Cytotoxic T cells

A

Develop into effector cells

Go looking for infected cells and kill them

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36
Q

B cells

A

Make antibodies of the IgM calss, then depending on the type of T cell help they receive undergo a class switch to other ab types like IgA, IgG, or IgE with the same original antigen specifity but different functions

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37
Q

Affinity maturation

A

Some cells will undergo modification and produce antibodies w/ increased affinity for the antigen which is called?

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38
Q

Plasma cells

A

B cells differentiate into these which reside in the bone marrow and secrete antibodies for the life of the individual

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39
Q

Neutralizing antibodies

A

Bind to the surface of microbes and prevent them from infecting cells

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40
Q

Opsonizing antibodies

A

Target microbes for uptake and destruction by phagocytic cells

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41
Q

Production of memory cells

A

Ultimate goal of vaccination

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42
Q

Leukocytes

A

Most numerous blood cell

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43
Q

Basophils

A

Least numerous blood cell

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44
Q

Neutrophils

A

Mediate early inflammatory responses

Segmented nucleus, cytoplasm contains granules which don’t stain well w/ dyes

Of the monocytic lineage

Short lived but act fast

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45
Q

Specific granules

A

Contain enzymes such as lysozyme, collagenase, and elastase

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46
Q

Azurophilic granules

A

Lysosomes containing other enzyme and microbicidal substances such as defensins and cathelicidins

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47
Q

Monocytes

A

Immature heterogenous circulating cells w/ little function

In blood

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48
Q

Macrophages

A

In tissue

Kill microbes, promote wound healing, resolve inflammatory responses

Guided by cytokines

Important for clearing apoptic cells

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49
Q

Apoptosis

A

Good death

Normal way cells turn over

Nuclear cleavage and breakdown of the cells into membrane encapsulated vesicles

These are recognized by macrophages and engulfed and destroyed w/ no leakage of cellular componenets

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50
Q

Necrosis

A

Bad death

Leads to disruption of the cell membrane and leakage of cellular contents (inflammatory)

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51
Q

Mast cells

A

Not normally in circulation

Reside beneath epithelial layers

Contain abundant granules filled w/ histamine and other vasoactive mediators

Coated w/ IgE Ab and are important in defense against helminths

Mediate allergic response

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52
Q

Basophils

A

Circulating blood granulocyte similar to mast cells

Coated w/ IgE, function in defense not certain

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53
Q

Eosinophils

A

Worm responses

Circulating blood granulocyte found lining mucosal sites

Granule enzymes are effective against parasitic infection but can also cause collateral damage to host cells

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54
Q

DC, B cells, and macrophages

A

What are the professional APCs?

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55
Q

Antigen presenting cells

A

Present antigen to T cells

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56
Q

Dendritic cells

A

Important for activating naive T cells

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57
Q

Macrophages

A

Important for antigen presentation to effector CD4 T cells at sites of infection

Causes the T cell to provide cytokine activation of the ? for enhanced microbial killing

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58
Q

B cells

A

Present antigen CD4 T cells in the lymph nodes and spleen and thereby receive help for antibody class switching

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59
Q

B and T lymphocytes

A

Cells of adaptive immunity

Express clonally distributed antigen receptors

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60
Q

T cells

A

Mature in thymus

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61
Q

CD3

A

Which marker is for all T cells?

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62
Q

ab Heterodimers except for gd T lymphocytes which use gd heterodimers

A

What do T cells use for antigen receptors?

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63
Q

CD4= II

CD8 = I

A

Which MHC complex do CD4+ use vs CD8+ T lymphocytes?

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64
Q

B cell differentiation

and

Macrophage activation

A

F ( CD4+ helper T lymphocytes)

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65
Q

Killing of infected cells and tumor cells

A

What is the function of CD8+ cytoxic T lymphocytes?

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66
Q

Suppresses other T cells so it regulates immune response

A

What is the function of Regulatory T cells?

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67
Q

Helper and cytotoxic functions

A

F ( gd T lymphocytes)

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68
Q

Ab production

A

F (B lymphocytes)

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69
Q

Killing of virus infected cells

A

F (NK cells)

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70
Q

Suppress/activate innate and adaptive immune response

A

F ( NKT cells)

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71
Q

NKT cells

A

What non T cell uses ab heterodimer as well?

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72
Q

Nk and NKT cells

A

Which cells use CD16 as markers?

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73
Q

Common lymphoid precursor > Bone marrow> blood lymph> lymph nodes and spleen> Mature

A

How do B lymphocytes mature?

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74
Q

Common lymphoid precursor> Thymus> Blood lymph> Mucosal and cutaneous lympoid tissues> Mature

A

How do T lymphocytes mature?

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75
Q

Nucleus: Naive
Activated: Cytoplasm

A

When does a lymphocyte have a lot of nucleus vs cytoplasm

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76
Q

Primary/central lymphoid organs

A

Where lymphocytes first express antigen receptors and attain their phenotypic and functional maturity
Thymus and bone marrow

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77
Q

Secondary/peripheral lymphoid organs

A

Where naive lymphocytes are activated and develop into effector cells
Spleen/lymph nodes

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78
Q

B lymphocytes, T lymphocytes, and NK cells

A

What does the common lymphoid progenitor give off?

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79
Q

Blood cells

A

What does the common myeloid progenitor give off?

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80
Q

Thymic dendritic cells

A

Present antigen to immature T cells and aid in their development

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81
Q

Thymocytes

A

Name for immature T cells

82
Q

Lymph node

A

Location of initiation of adaptive immune responses to lymph borne antigens

83
Q

Blood stream

A

How do naive lymphocytes enter the lymph node?

84
Q

Dendritic cells thru afferent lymphatic vessels

A

How does antigen enter lymph nodese?

85
Q
T= CCR7
B= CXCR5
A

What receptors do naive T and B cells use?

86
Q

Spleen

A

Location where aging and damaged blood cells and opsonized microbes are removed and destroyed

87
Q

Homing

A

Process of directing cells to their functional sites

88
Q

Recruitment

A

Movement of cells from the circulation into sites of infection or injury.
Inflammation

89
Q

Adhesion of the leukocytes to the enodthelial lining

A

What does leukocyte recruitment depend on?

90
Q

Selectins

A

Mediates the initial step of Low affinity binding of leukocytes to endothelium

91
Q

L selectin

A

Expressed on lymphocyte

Helps naive T and B cells get out of vasculature at site of lymph node.

92
Q

E and P selectins

A

On endothelial surface/side

Expression is increased by TNF and IL-1

93
Q

Integrins

A

Mediate increased binding of cells to endothelium
Can be on leukocyte or endothelium

Expression increased by cytokine

Leukocyte ones start as low affinity and become high affinity upon activation

94
Q

Partially activated when encounters receptor

A

How are integrins activated?

95
Q

Chemokines

A

Tell cells where to go

Structurally homologous cytokines that stimulate leukocyte movement bot in homing and recruitment

96
Q

CXC, CC, C, CXXXC

A

What are the classes of chemokines?

97
Q

CXCL8 is a chemokine

CXCR1 is a receptor

A

What is CXCL8 vs. CXCR1

98
Q

Chemokines and cytokines flag down neutrophils causing a rolling movement. The flags are most concentrated at the center of the infection. Neutrophils slow down. At some point high affinity integrin is expressed which causes it to stop and flatten and crawl until it finds a spot where it can get b/t 2 endothelial cells and get to vasculature

A

How are leukocytes recruited into tissues?

99
Q

L-selectin, CCR7, LFA-1

A

What are the naive T cell homing receptors?

100
Q

E and P selectin ligand, CXCR3, CCR5, LFA 1, and VLA 4

A

What are the activated T cell receptors?

101
Q

L selectin: L selectin ligand
CCR7: CCL19 or CCL21

LFA-1: ICAM-1

A

What are the ligands for the receptors of the naive T cells?
L selectin:

CCR7:

LFA-1:

102
Q

E and P selection ligand: E of P selectin
CXCR3: CXCL10

CCR5: CCL4

LFA-1:ICAM or VCAM

A

What are the Activated T cell ligands?
E and P selectin ligand:

CXCR3

CCR5

LFA-1:

103
Q

L selectin

A

Responsible for initial weak adhesion of naive T cell to high endothelial venule in lymph node

104
Q

LFA w/ ICAM

A

Stable arrest on high endothelial venule in lymph node

105
Q

E and P selectin ligand

A

Responsible for initial weak adhesion of effector and memory T cells to cytokine activated endothelium at site of infection

106
Q

Activate integrins and chemokines

A

What do CCXR and CCXL’s do?

107
Q

Innate immunity

A

First line of defense against infection
Exists prior to encounter w/ microbe

Rapid response

Effective defense against most microbes, but pathogenic microbes can evade some defenses

Directs development of adaptive immunity

108
Q

Not known consciously> Inflammation> Naive B and T cells (adaptive immune response)>Clonal expansion

A

Phases of initial immune response

109
Q

Pathogens

A

Microbes that can cause disease

110
Q

Virulence

A

Degree to which a pathogen can cause disease

111
Q

Host sees it is being invaded

A

First step in innate immunity

112
Q

Sentinel cells

A

Have surface receptors for molecules that are normally expressed by microbes, but never in higher animals
Recognize the presence of microbes

Live in tissues, not circulation

113
Q

PAMPS (Pathogen-associated molecular patterns)

A

Microbial substances that stimulate innate immunity

Recognized by pattern recognition receptors

114
Q

Germline, limited diversity

A

Where are receptors of innate immunity made?

115
Q

Somatic recombination of gene segments, greater diversity

A

Where are receptors of adaptive immunity made?

116
Q

Innate: Nonclonal
Adaptive: Clonal

A

Is innate/adaptive clonal/nonclonal

117
Q

Clonal

A

Copies of all lymphocytes w/ distinct specificities express different receptors

118
Q

Adaptive

A

Does innate/adaptive have autoimmunity?

119
Q

DAMPs or Damage associated molecular patterns

A

Endogenous molecules that are produced or released from damaged and dying cells
Recognized by immune system

Not typically released by apoptotic cells

120
Q

LPS

A

What is the main thing that gets recognized on gram negative bacteria?

121
Q

PRR recognizees teichoic acid, surface protein, and lipoteichoic acid

A

What recognizes what on gram positive bacteria?

122
Q

TLR4 with LPS

A

What is the big TLR for gram negative bacteria?

123
Q
  1. TLR1 and 2
  2. TLR2
  3. TLR5
  4. TLR 2 and 6
A

What are the TLRs for
1. Bacterial lipopeptides

  1. Bacterial peptidoglycan
  2. Bacterial flagellan
  3. Bacterial lipopeptides
124
Q

TLR 7 and 8

A

Which TLRs recognize ssDNA in the endosome?

125
Q

MyD88 except TLR3 (thru TRP)

A

All TLRs go through _______ except ________ which signals thru _____

126
Q

TLR3 w/ TRP

A

What signals interferon response genes to be activated and starts an antiviral response and signals to make type I interferons?

127
Q

Macrophage mannose receptor

A

Binds terminal mannose and fucose residues of glycoproteins and glycolipids

128
Q

Scavenger receptors

A

Bind to acetylated low density lipoproteins. Sialated in mammals but not microbes

129
Q

N-formylmethionyl receptors

A

All bacterial proteins and few mammalian proteins are initiated w/ N-formylmethionine

130
Q

Defensins

A

Small cationic peptides produced by epithelial cells, neutrophils, NK cells and CTL. Directly toxic to microbesand activate inflammatory cells
Have antibiotic effects

131
Q

Cathelicidins

A

Produced by neutrophils and epithelial cells

Produced as a precursor, cleavage stimulated by cutokines. Can bind and neutralize LPS

132
Q

CD14

A

What is part of TLR4 binding to LPS?

133
Q

Cytoskeletal changes > Migration into tissues

A

What does activation of the seven a-helical transmembrance receptors cause?

134
Q

Killing of microbes

A

What does activation of TLRs or mannose receptors lead to?

135
Q

NK cells

A

Cells of the lymphocyte lineage that recognize stressed or infected cells and directly kill them

136
Q

Secreting IL-12 to NK cells and NK cells activating macrophage by secreting IF-y

A

What gives macrophages the extra added capability of killing microbes?

137
Q
  1. Kill things

2. Activate macrophages

A

2 functions of NK cells

138
Q

2 receptors: activating and inhibitory (dominant)
If the inhibitory receptor is engaged, or the Class I MHC complex is present, the cell will not be killed

If the inhibitory complex is not engaged or if the MHC is inhibited, the cell will be killed.

A

How do NK cells kill things?

139
Q
  1. B-1 cells
  2. Epithelial y:S cells
  3. NKT cells
A

What are the innate like lymphocytes?

140
Q

Make natural Ab

A

F (B-1 cells)

141
Q

Produce cytokines rapidly

A

F( Epithelial y:S and NKT cells)

142
Q

MHC class IB

A

What are the ligands of the Epithelial y:S cells?

143
Q

Lipids bond to CD1

A

What is the ligand and what is it bound to of the NKT cells?

144
Q

No

A

Can innate like lymphocytes be boosted?

145
Q

B-1 cells

A

Present in peritoneal cavity and spleen
Limited receptor diversity

Specific for polysaccharide and lipid antigens

Produced w/o evidence of infection

Mainly IgM

146
Q

T-independent antigen

A

What makes IgM?

147
Q

Binds to microbes and activates complement

A

What does IgM do?

148
Q

Opsonins

A

Soluble molecules that bind to the surface of microbes and induce their uptake by phagocytic cells thru specific receptors on the cell for the opsonin molecule

149
Q

Complement pathway

A

Series of plasma proteins work together to opsonize microbes, promote recruitment of phagocytes to the site of infection and in some cases directly kill microbes. Can activate inflammatory response

150
Q
  1. Classical- Ab bound to antigen
  2. MB-Lectin
  3. Alternative-Complement can bind directly to microbe
A

3 ways to start complement pathway

151
Q
  1. Recruitment of inflammatory cells
  2. Opsonizations of pathogens
  3. Killing of pathogens
A

3 outcomes of complement pathway

152
Q

Inflammation

A

What do C3A and C5A cause?

153
Q
  1. C3 is cleaved into C3A and C3B
  2. C3A(inflammatory) is released, C3B stays w/ the microbe (becomes C5)
  3. C5 is cleaved into C5A (inflammatory) and C5B (which stays w/ microbe)
  4. If this keeps going, causes MAC pore which causes osmotic lysis
A

What happens after complement is activated?

154
Q
  1. Opsonization and phagocytosis
  2. Stim of inflammatory rxns
  3. Complement mediated cytolysis
A

3 functions of complement

155
Q

Inflammatory response

A

Major outcome of innate immune activation is the induction of acute inflammation which is the accumulation of leukocytes, plasma proteins, and fluid from the blood at an extravascular site of tissue injury or infection

156
Q
  1. IL-1
  2. TNF
  3. IL-6
  4. CXCL8
  5. IL-12
A

First cytokines made in innate immunity (5)

157
Q

Fever, shock, mobilization

A

What are the systemic effects of cytokines?

158
Q

TNF

A

What cytokine is known to cause shock?

159
Q

IL-1 and TNF

A

Which cytokines activate vascular endothelium?

160
Q

IL-12

A

Which cytokine activates NK cells

161
Q

Radicals

A

How do INOS and oxidase kill?

162
Q

Killing microbes
Inflammation enhanced adaptive immunity

Tissue remodeling

A

What are the effector functions of activated macrophages?

163
Q

ROS and Nitric Oxide

A

Which molecules produced in activated macrophages kill microbes?

164
Q

TNF

A

Responsible for negative sytemic effects
Shock

Can’t be stopped once it starts

Causes low output in the heart, thrombus, increased permeability in blood vessels, and Insulin resistance

165
Q

Cause fever and leukocyte production

IL 6 and IL 1 also cause acute phase proteins which are opsonins

A

What do TNF, IL-1, and IL-6 do systemically that is positive?

166
Q

Antiviral response

A

Protection agains viruses is primarily an adaptive immunity concern, but innate plays a role too

167
Q
  1. Inhibition of viral protein synthesis
  2. Degradation of viral RNA
  3. Inhibition of viral gene expression and virion assembly
A

What are the 3 parts of the antiviral state? The three actions of type I IFNs?

168
Q

Type I: a and B

Type II: gamma

A

What are Type I vs Type II IFNs?

169
Q

Ab

A

Circulating substance w/ in the serum of animals immunized w/ an attenuated form of diphtheria toxin that could provide protection against challenge w/ the virulent organism
AKA antitoxin/Ig

170
Q

Antigen

A

Substance Ab recognize

171
Q

Membrane bound and soluble

A

2 forms of Ab

172
Q

Membrane bound Ab

A

Function as B cell receptor

173
Q

Soluble Ab

A

From antigen stimulated B cells (plasma cells)

174
Q

Serology

A

[] of Ab for an antigen determined via serial dilutions

Last binding observed = titer

175
Q

Neutralization
Activation of complement system

Opsonization of pathogens

Ab-dependent cell-mediated cytotoxicity

Ab-mediated mast cell activation

A

(5)Effector functions

176
Q

Fc Region

A

Part of Ab
Has effector function

Non-antigen binding portion

Few variations b/t Ab classes

177
Q

Fab region

A

Part of Ab
Antigen binding site

Highly variable

Distinguish Abs made by one clone of B cell vs another clone

178
Q

Gamma > Ig

A

Which globulin makes up the biggest chunk of globulins?

179
Q

Monoclonal

A

Single clone
Steep peak

Indicates cancer

180
Q

CDRs (Complementarity-determining Regions) 1, 2, and 3

A

Most of the Ab variability is contained w/ in 3 short regions called the hypervariable region

181
Q

CDR3

A

Which CDR has most contact w/ antigen?

182
Q

K and gamma

A

2 types of light chains that are functionally the same?

183
Q

IgA: Mucosal immunity
IgE: Defense agains Parasites and hypersensitive

IgG1: Opsonization complement activation

IgM: 1st responder

A

Function of
IgA

IgE

IgG1

IgM

184
Q
IgM+ = Early infection
IgG+ = Later stage infection
A

When you test cats for toxo, what do the results mean?

185
Q

Secreted Ab

A

Short tail Ab

Hydrophilic

186
Q

Membrane bound

A

Transmembrane region

Hydrophobic

187
Q

Monoclonal Ab

A

Specific for specified single epitrope on antigen
Fuse B cells from immunized animal w/ myeloma cell line

Purpose: Diagnosis of infectious and systemic diseases

Targeting cells and molecules

Detection of tumors

188
Q

Rearrangement and expression of H and L chains

A

What accompanies maturation of B cells from bone marrow progenitors?

189
Q

Antigen

A

Any substance that can by sepcifically bound by an ab or T cell receptor

190
Q

Epitope

A

Portion of antigen bound by Ab or TCR

191
Q

Hapten

A

Small chemical too small to be immunogenic (to induce an Ab response)
Won’t stim immune system

192
Q

Carrier

A

Protein or polysaccharide that is conjugated to hapten to act as an immunogen (induce Ab response)

193
Q

Hapten carrier complex

A

Induces Ab response to hapten

Ex: vaccines

194
Q

Determinant

A

Epitrope

Portion of macromolecule Ab binds to

195
Q

Linear determinant

A

On external surface or inaccessible unless denatured
Adjacent AA recognized

AA are together

196
Q

Conformational determinant

A

AA residues not in sequence but become spatially positioned next to each other in the folded protein

197
Q

Neoantigenic determinants

A

Modification of proteins by glycolsylation, phosphorylation, ubiquitination, acetylation, and proteolysis
Production of new epitopes not present before modification

198
Q

Affinity

A

Strength of binding b/t single epitope and ab binding site

199
Q

Avidity

A

Cumulative strenght of attachment of all Ab binding sites to epitopes

200
Q

Change in heavy chain of constant region

A

How do isotypes switch?