Separation of alveolar surfactant into subtypes - A comparison of methods

被引:17
作者
Gross, NJ
Kellam, M
Young, J
Krishnasamy, S
Dhand, R
机构
[1] Hines Vet Affairs Hosp, Med Serv, Hines, IL 60141 USA
[2] Hines Vet Affairs Hosp, Res Serv, Hines, IL 60141 USA
[3] Loyola Univ, Stritch Sch Med, Dept Mol Biochem, Hines, IL USA
[4] Loyola Univ, Stritch Sch Med, Dept Med, Hines, IL USA
关键词
D O I
10.1164/ajrccm.162.2.9908081
中图分类号
R4 [临床医学];
学科分类号
1002 ; 100602 ;
摘要
Alveolar surfactant is known to exist in several morphologic forms or subtypes which have been separated from bronchoalveolar lavage fluid (BAL) by two types of methods-differential centrifugation (DC) and equilibrium buoyant density gradient centrifugation (EBDC). DC separates BAL into large aggregates (LA) and small aggregates (SA); EBDC separates BAL into three peaks called ultraheavy (UH), heavy (H), and light (L). We compared these two separation methods by subjecting replicates of the same pools of BALF from groups of mice to DC and EBDC in parallel assays. We found that each method was highly internally consistent, but that the amount of phospholipid in the LA fraction of DC was consistently and substantially less (by 33 to 43%) than that found in the UH + H fractions of EBDC, This appeared to be due to failure of DC to sediment all of the phospholipid that banded as UH or H in EBDC despite adjustments in the time and g-force of DC. In experiments where differentially labeled purified H and L subtypes were subjected to DC over a wide range of g-force and time conditions, cross-contamination of the DC pellet and supernatant with heterologous subtypes was always present (4 to 33% cross-contamination), Addition of extraneous serum proteins to the BAL, as a model of lung damage, resulted in further inconsistencies in DC but not EBDC, Investigators may wish to bear these considerations in mind when planning or interpreting the results of experiments bearing on surfactant subtype analysis.
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页码:617 / 622
页数:6
相关论文
共 14 条
  • [1] SP-A, SP-B, AND SP-C IN SURFACTANT SUBTYPES AROUND BIRTH - REEXAMINATION OF ALVEOLAR LIFE-CYCLE OF SURFACTANT
    BARITUSSIO, A
    ALBERTI, A
    QUAGLINO, D
    PETTENAZZO, A
    DALZOPPO, D
    SARTORI, L
    PASQUALIRONCHETTI, I
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1994, 266 (04): : L436 - L447
  • [2] Folch J., 1957, J BIOL CHEM, V226, P498
  • [3] SURFACTANT SUBTYPES OF MICE - METABOLIC RELATIONSHIPS AND CONVERSION INVITRO
    GROSS, NJ
    NARINE, KR
    [J]. JOURNAL OF APPLIED PHYSIOLOGY, 1989, 67 (01) : 414 - 421
  • [4] SURFACTANT SUBTYPES IN MICE - CHARACTERIZATION AND QUANTITATION
    GROSS, NJ
    NARINE, KR
    [J]. JOURNAL OF APPLIED PHYSIOLOGY, 1989, 66 (01) : 342 - 349
  • [5] INVITRO CONVERSION OF SURFACTANT SUBTYPES IS ALTERED IN ALVEOLAR SURFACTANT ISOLATED FROM INJURED LUNGS
    HIGUCHI, R
    LEWIS, J
    IKEGAMI, M
    [J]. AMERICAN REVIEW OF RESPIRATORY DISEASE, 1992, 145 (06): : 1416 - 1420
  • [6] Ventilation strategies affect surfactant aggregate conversion in acute lung injury
    Ito, Y
    Veldhuizen, RAW
    Yao, LJ
    McCaig, LA
    Bartlett, AJ
    Lewis, JF
    [J]. AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, 1997, 155 (02) : 493 - 499
  • [7] SUBFRACTIONATION OF LUNG SURFACTANT - IMPLICATIONS FOR METABOLISM AND SURFACE-ACTIVITY
    MAGOON, MW
    WRIGHT, JR
    BARITUSSIO, A
    WILLIAMS, MC
    GOERKE, J
    BENSON, BJ
    HAMILTON, RL
    CLEMENTS, JA
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1983, 750 (01) : 18 - 31
  • [8] Characterization of surfactant subtypes of beractant and a synthetic peptide containing surfactant KL4 following surface area cycling and addition of fibrinogen
    Manalo, E
    Merritt, TA
    Amirkhanian, JD
    Kheiter, A
    [J]. LUNG, 1997, 175 (04) : 225 - 233
  • [9] MOUSE ALVEOLAR SURFACTANT - CHARACTERIZATION OF SUBTYPES PREPARED BY DIFFERENTIAL CENTRIFUGATION
    OULTON, M
    MACDONALD, J
    JANIGAN, DT
    FAULKNER, GT
    [J]. LIPIDS, 1993, 28 (08) : 715 - 720
  • [10] Convertase activity in alveolar surfactant and lamellar bodies in fetal, newborn, and adult rabbits
    Oulton, M
    Edwards, E
    Handa, K
    [J]. JOURNAL OF APPLIED PHYSIOLOGY, 1999, 86 (01) : 71 - 77