Differential allergy responses to Metarhizium anisopliae fungal component extracts in BALB/c mice

Figures & data

Figure 1.  BALF total and differential cell counts. Data shown represent mean ± SE. (§) indicates significant differences compared to HBSS, BSA, HB/MY, HB/IN; or (≠) compared to HBSS, BSA, HB/MY only. Letter (A = MACA, C = CON, I = IND, M = MYC) identifies differences among treatments: at p < 0.05. n = 6.

Figure 2.  (A) Serum and BALF total IgE. (B) BALF total IgA. (C) Antigen-specific IgE quantified in functional assay by measurement of mast cell mediator (β-hexosaminidase) release and presented as percent of total mediator release. Data shown represent mean ± SE. Significantly elevated compared to (§) all controls; (#) to other treatments; (≠) HBSS and BSA at p < 0.05. n = 6.

Figure 3.  (A) Immediate respiratory physiological responses averaged for 1 hr immediately following IA extract exposure. (B) Respiratory responses to increasing concentrations of methacholine aerosol at D1 and D3 after final IA exposure. Data shown represent mean ± SE. Significantly elevated compared to (#) HBSS and/or BSA; (§) all controls; (≠) all treatments; other treatments: C = CON, I = IND, M = MYC, A = MACA at p < 0.05. n = 6.

Table 1.  Summary of incidence and severity of mouse lung pathology.

	Incidence^a
	(Group Severity)^b
	MACA	MYC	CON	IND	HB/MY	HB/IN	HBSS	BSA
Bronchial Epithelial Hypertrophy	6/6	7/7	5/6	6/6	0/6	0/6	0/6	0/6
	(2.0)	(2.1)	(1.5)	(2.0)
Peribronchial Mononuclear Cell Infiltration (predominately lymphocytes)	6/6	7/7	6/6	6/6	6/6	1/6	1/6	2/6
	(3.3)	(3.6)	(2.8)	(2.8)	(2.2)	(0.3)	(0.2)	(0.7)
Peribronchial Polymorphonuclear Cells (predominately neutrophils)	6/6	7/7	4/6	6/6	1/6	0/6	0/6	1/6
	(2.2)	(1.9)	(0.7)	(1.7)	(0.3)			(0.2)
Perivascular Mononuclear Cell Infiltration	6/6	7/7	6/6	6/6	6/6	6/6	1/6	6/6
	(2.3)	(3.3)	(2.5)	(2.0)	(1.3)	(1.0)	(0.2)	(1.2)
Perivascular PMN Cell Infiltration	6/6	7/7	4/6	6/6	1/6	2/6	0/6	3/6
	(2.2)	(2.0)	(0.8)	(2.0)	(0.3)	(0.3)		(0.5)
Alveolitis, Focal	1/6	3/7	0/6	0/6	2/6	1/6	1/6	0/6
	(0.2)	(0.7)			(0.5)	(0.2)

^aIncidence indices were determined by the number of mice with the pathological feature over the number of mice examined.

^bGroup Severity indices were determined by dividing the sum of the numerical score for a particular lesion by the number of animals examined.

Figure 4.  (A) The neurotrophins NGF, NT-3 and NT-4 in BALF were assayed by ELISA. The limit of detection was 15.6 pg/ml for NGF, 4.7 pg/ml for both NT-3, and NT-4. Data shown represent mean ± SE. Significantly elevated compared to (§) all controls; (≠) HBSS and/or BSA; other extracts: C = CON, I = IND, M = MYC, A = MACA at p < 0.05. n = 6. (B) Protease activity is shown as percent of trypsin control.

Figure 5  A Western blot of MACA and M. anisopliae component extracts were probed with mouse anti-MACA hyperimmune serum and HRP-labeled rat anti-mouse IgE to identify extract proteins that bind IgE.

Table 2.  Response summary table.

	BALF Total		Serum Total	Specific IgE	Neurotrophins			Cell Counts		Airway Responses			Protease Activity
	IgA	IgE	IgE	% Tot Rel	NGF	NT4	NT3	Total Cells	Eos	PenH/D0	Mch Chal 64 D1	Mch Chal 64 D3	% of Trypsin
MACA	3022.8	100.5	14437	61.8	337.3	398.9	1775.2	60.6	35.7	427.4	1475.1	976.1	481.5
MYC	7848.8	284.5	37395	66.6	1470.5	1660.6	3563.6	72.9	36.9	209.3	1260.9	962.6	42.8
CON	2169.6	29.5	8281.9	18.6	857.7	1022.2	2266.3	42.5	17.1	339.8	1217.2	852.0	36.7
IND	1930.4	28.8	12239	22.9	232.6	162.5	678.2	36.8	17.1	600.5	1942.8	1451	477.8
HB/MY	604.3	12.9	1682.6	0.4	101.3	59.7	266.7	5.4	0	8.1	920.7	749.7	nd
HB/IN	536	24.3	2197.6	− 0.4	332.1	442.8	265.5	26.9	0.9	5.6	969.3	671.9	nd
HBSS	115.7	12.5	2500.5	0	86.3	22.9	82.4	5.4	0	7.5	560.6	479.0	nd
BSA	126	12.5	1887.9	− 1.1	110.8	16.2	74.3	4.9	0	8.6	550.5	478.1	nd

Highest magnitude of response for each endpoint is shaded