Immunochemical Studies on the Antigens of Bordetella Pertussis

Abstract

Sonic extracts of B. pertussis were fractionated by chromatography employing DEAE-cellulose. Mouse protective antigen (MPA) was absorbed to the column and was found to be eluted over a range of salt concentrations from 0.05 to 0.35 M NaCl in dilute phosphate buffer, pH 7.2. The MPA was freed from nucleic acid impurities since these remained fixed to the column and could only be eluted with salt concentrations of 0.5 to 2.0 molarity. Some impurities of protein nature passed directly through the column. The measurable precipitating antigens associated with the crude extract were fractionated into two groups by the DEAE-cellulose. Dilute buffer elutions removed material that developed several cathodic bands by immunoelectrophoresis. More concentrated salt buffers eluted materials that exhibited one anodic band by immunoelectrophoresis against the whole B. pertussis antiserum. The measurement of significant potencies of MPA in the absence of either cathodic or anodic precipitinogens suggests that the latter represent impurities which may or may not have a relation to other known biological activities associated with B. pertussis (e.g., toxin).Certain discrepancies between mouse protective antigen and histamine-sensitizing factor based on the lack of proportionality of the activities from fraction to fraction suggest that their chemical natures are dissimilar.

Histamine-sensitizing factors from microbial agents, with special reference to Bordetella pertussis.

“…The work of Nagel (170) also shows that HSF and protective antigen are separable. These results support previous claims that HSF and protective antigen are two different substances (43,74,210,227).…”

Inheritance of Responsiveness to Pertussis HSF in Mice

“…The pertussis component producing this effect was designated 'histaminesensitizing factor' (HSF) by M a it l a n d , C o h n and M a c d o n a l d [10] and appears to occur in phase 1 strains of B. pertussis and in no other bacterium except possibly Brucella abortus [11]. Partially purified HSF has been obtained by M u n o z and H est e k in [14], N iw a [18] and G r if f it h s and M a so n [5] but its chemical nature is not yet known. However, current evidence points to HSF being located in the bacterial cell wall [12] and containing protein in its make up [12,21,29], Some investigators [9,12,21] have suggested that HSF may be the same as the protective antigen (PA) of B.pertussis that is responsible for immunizing children against whooping cough, but there are also contrary opinions [3,17,23].…”

Purification of Histamine Sensitizing Factor of Bordetella Pertussis

“…A peak of lethal toxicity exuited in fraction 3. These data demonstrate that 11SF as well as lethal activities can not bc tefer to riboso" e particle itself.…”