Surfactant Composition and Function

Joanna Floros, Ph.D

Evan Pugh Professor, Departments of Cellular and Molecular Physiology, Pediatrics, and Obstetrics and Gynecology. The Pennsylvania State University, College of Medicine, Hershey, PA, U.S.A

8th World Hellenic Biomedical Congress

Surfactant
1. Characteristics of Surfactant What is surfactant? What is its composition? Where is it made? What is its structure?

2. Function of surfactant

What is Pulmonary Surfactant and What is its Composition?

Pulmonary surfactant biochemically, is a lipoprotein complex that lines the internal surface of the alveoli. MAJOR CONSTITUENTS OF PULMONARY SURFACTANT Unsaturated PC

PC (DSPC)

PE
16:/16:0

Protein
Other lipids

SP-A SP-B SP-C SP-D

Other Proteins

Cholesterol

PG PI

Surfactant

Proteins

Fishman 2-19

SURFACTANT HETEROGENEITY AND LIFE CYCLE

LAMELLAR BODIES TUBULAR MYELIN

SP-A SURFACE FILM

LARGE AGGREGATES LB, TM, SP-A

SMALL AGGREGATES

HYPOPHASE FLUID (SUPERNATANT)

Surfactant
1. Characteristics of Surfactant What is surfactant? What is its composition? Where is it made? What is its structure?

2. Function of surfactant and/or surfactant components Lowering of surface tension. Innate host defense. Initiation of parturition (SP-A, a hormone?)

Functions of surfactant
Surface tension

Stabilizes alveoli and prevents alveolar (lung) collapse with each exhalation by lowering the surface tension at the air-liquid surface. Host defense Surfactant components play a role in host defense and protect the lung from injuries or infections caused by inhaled microorganisms, particles, toxins, other. Initiation of labor One of the protein component of surfactant (SP-A) may serve as a hormone in parturition.

Air-Liquid Interface Without surfactant With surfactant

Surfactant

J. Kending

MAJOR CONSTITUENTS OF PULMONARY SURFACTANT

Unsaturated PC PC (DSPC) PE
16:0/16:0

Protein
Other lipids

SP-A SP-B SP-C SP-D

Other Proteins

Cholesterol

PG PI

Surfactant

Proteins

Functions of surfactant
Surface tension

Stabilizes alveoli and prevents alveolar (lung) collapse with each exhalation by lowering the surface tension at the airliquid surface. Host defense Surfactant components play a role in host defense and protect the lung from injuries or infections caused by inhaled microorganisms, particles, toxins, other.
Initiation of labor One of the protein component of surfactant (SP-A) may serve as a hormone in parturition.

SURFACTANT REGULATION OF IMMUNE CELLS

PE SM CHOL SP-A

SP-A Stimulation

LIPID Inhibition
STIMULATION

PI PG PC

INHIBITION

Oxidant Production Splenocyte Proliferation Cell Surface Markers Cytokine Production NF-kB Activation Fibroblast Apoptosis Collagen and Collagenase Production

PI PG PC

PE SM CHOL SP-A

STIMULATION

INHIBITION

SP-A FUNCTION
Surfactant-related SP-A Host defense-related
Innate Immunity

Innate
800 700
TUBULAR MYELIN

Adaptive
500 present

600 myr

Regulation of inflammatory processes

LARGE AGGREGATES

Cell Surface Markers; Cytokine Production; NF-kB Activation; Collagen and Collagenase Production

Human SP-A Genes, and Variants

Genes

SP-A2

P
1A0 6A2

SP-A1

Variants

The above variants have been shown to exhibit qualitative and quantitative differences. In addition environmental insults, such as ozone, appear to differentially alter their functional activity and thus contribute to individual differences in disease susceptibility.

Differences exist among SP-A variants in their ability to enhance phagocytosis of bacteria by alveolar macrophages in the presence or absence of environmental factors.

EXPERIMENTAL APPROACH
agar plate

suspension of bacteria

ozone BAL

wash off free bacteria

insect cells or mammalian cells SP-A phagocytosis slide preparation

light microscopy analysis

rat

alveolar macrophages

RESULTS
Microscopic view of live P. aeruginosa, phagocytized by rat alveolar macrophages in the presence or absence of human SP-A No SP-A SP-A

SP-A2 products exhibit higher phagocytic ability than SP-A1

Mammalian expressed variants

500

*** * *
hSP-A SP-A2

Phagocytic index (%)

400

300

**
SP-A1
6A2 6A4

200 1A0 1A

100

1.56 mg/ml

n=5

* SP-A2 vs. SP-A1 ;

** 6A2 vs. 6A4 ; *** hSP-A vs. 6A4 (mammalian); (hSP-A: 25 g/ml) Bacteria: P.aeruginosa

hSP-A vs. SP-A variants (insect)

Phagocytic index: % of bacteria-positive AMs the average # of bacteria per bacteria-positive AMs Mikerov et al

Ability of SP-A variants to enhance phagocytosis by alveolar macrophages following exposure to ozone.

Unexposed

Ozone-exposed 1A0 6A4 1A0/6A4

1A0 > 6A4 = 1A0/6A4 >

(SP-A2) (SP-A1) (SP-A2/SP-A1)

Functions of surfactant
Surface tension Stabilizes alveoli and prevents alveolar (lung) collapse with each exhalation by lowering the surface tension at the airliquid surface.

Host defense Surfactant components play a role in host defense and protect the lung from injuries or infections caused by inhaled microorganisms, particles, toxins, other. Initiation of labor One of the protein component of surfactant (SP-A) may serve as a hormone in parturition.

Fetal SP-A initiates labor

Increased levels of SP-A
activate Fetal macrophages

Surfactant lipids
(Arachidonate derivatives)

Increased levels of proinflammatory cytokines (i.e. IL-b1) activate NF-kB

Prostaglandin synthesis

Increase expression of genes that cause myometrial contractibility

P65 antagonizes progesterone receptor activation of genes that modulate uterine quiescence.

Summary
Pulmonary surfactant, a lipoprotein complex, is essential throughout life for normal lung function, host defense, and other functions. Derangement in surfactant either through alterations in composition or modification (i.e. oxidation) can negatively affect normal lung function and host defense. Genetic differences in surfactant components may contribute to differences in disease risk among individuals.