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Transfer Factor Therapy
Revisited
J. Kelly Smith, M.D., F.A.C.P.
DISCLAIMER
NEITHER THE PUBLISHER NOR THE AUTHORS
ASSUME ANY LIABILITY FOR ANY INJURY AND
OR DAMAGE TO PERSONS OR PROPERTY
ARISING FROM THIS WEBSITE AND ITS
CONTENT.
Ilya Metchnikoff
Phagocytosis theory
“eating
g to eat”
Ilya Metchnikoff
Phagocytosis theory
“eating to defend”
Hans Ernst Buchner
Humoral theory of immunity
George Nuttel
Karl Landsteiner & Merrill
Chase
Cell--mediated immunity
Cell
Proc Soc Exp Biol Med 1942;49:688-690
Transfer of delayed cutaneous
hypersensitivity
A. Picryl chloride + MTb
B. DCH – to DCH +
C. Peritoneal exudates out
D. Washed peritoneal exudates in
E. DCH – to DCH +
Sherwood Lawrence
Transfer factor
Proc Soc Exp Biol Med 1949;71:516-522
She ood Lawrence
Sherwood
La ence
Lymphocytes or their lysates
DCH transfer specific to donor
Rapid
R id in
i onsett
Long duration
Not inactivated by trypsin, DNase or RNase
Low molecular weight “informational” or
“ ti t ” molecule
“activator”
l
l
Gene de
de--repressor?
Othe s
Others
Colostrum & cloned lymphocytes
DCH transfer both specific & nonspecific
>200 highly polarized, hydrophilic, low
molecular weight peptides
Functions
F nctions ac
across
oss species
Well tolerated
tole ated & stable
Non-immunogenic
NonNo viruses > 10,000 mw
Mild pain at site of injection
Readily sterilized
Can store in lyophilized state > 5 years
Occassional flare of illness
Similarities to thymosins
Thymosin--α
Thymosin




Pl i t hi peptide
Pleiotrophic
tid
Activates moDC and pDC subsets
Promotes T & NK cell maturation
Stimulates cytokine production and CTLCTLmediated
di t d cytotoxicity
t t i it
Simila ities to thymosins
Similarities
th mosins
Prothymosin--α
Prothymosin



A histone H1
H1--binding polypeptide
Synergizes with CREB binding protein to
stimulate AP1
AP1-- and NF
NF--KB-dependent
transcription
i i by
b chromatin
h
i remodeling
d li
Immune stimulation is mediated primarily by
i d ti off monocyteinduction
monocyte
t -derived
d i d dendritic
d d iti cellll
maturation and by monocyte activation
Dend itic cells
Dendritic
Immature
Mature
Dend itic cells
Dendritic








Premier antigenantigen-presenting cells (MHC I & II)
Activate selfself-replicating (IL(IL-2 +) cytotoxic T
cells
Activate thymocytes
Activate T helper cells
Activate macrophages
Promote TH1 differentiation
Kill viruses (plasmacytoid DCs)
Detect PAMPs (TLRs)
Mac ophages
Macrophages
 Antigen
presentation (MHC II)
 Immune response amplification
 Inflammation
I fl
ti
 Pathogen ingestion/killing
 Apoptotic T cell clearance
 Chronic graft rejection
Clinical Studies
Patients
Candidiasis & congenital thymic disorders
Congenitall CMV infection
f
Prosthetic valve candida endocarditis
Chronic active hepatitis
Disseminated molluscum contagiosum
Recalcitrant malignancies
My first medical record (circa 28
28,000
000 years b
b.c.)
c)
Methods
TF prepared from leukocyte dialysates (DLE)
(<10,000 molecular weight)
DCH testing with candida, SK
SK--SD, trichophyton,
mumps antigens +/
+/-- DNCB
Standard assays
y were used to measure T & B
cells, immunoglobulins, ABO isoagglutinins, HMP
shunt activity, complement, leukocyte chemotaxis,
migration inhibition
hb
factor
f
(MIF),
(
) mitogenic
responses, and Fcγ
FcγRs.
Methods
T rosette assay: an early marker for CD3+ T cell
activation



Both CD4+ & CD8+ T cells & thymocytes
Correlates with DCH to specific antigen, allogeneic cell
cytotoxicity, and B cell activation
Rosetting populations increase with mitogenic
stimulation (PHA, ConA)
Methods
Migration Inhibition Factor (MIF)





Multifunction pro
pro--inflammatory cytokine produced by TTlymphocytes & macrophages
Suppresses antianti-inflammatory effects of glucocorticoids
Increases endothelial leukocyte recruitment and adhesion
Direct chemokine effects
Induces DC maturation and IL
IL--12 production
Congenital thymic disorders
Autoimmune polyendocrinopathy
syndrome
y
type
yp 1
APS--type
APS
t pe 1
M t ti
Mutations
iin ttranscriptional
i ti
l regulator
l t and
d
proaptotic factor AIRE in the thymus & in
monocyte--derived dendritic cells (moDCs)
monocyte



Impaired clonal deletion of selfself-reactive thymocytes.
Autoimmune destruction of adrenal & parathyroid
glands
Tolerance to candida infection
APS--type
APS
t pe 1
Positive selection
MHC–self peptide recognition = promoted
Thymic epithelial cells
N
Negative
ti selection
l ti
MHC-self peptide recognition = fail (98%)
APS-1 = g
graduate
Thymic dendritic cells
K F.
K.
F
CMC at 3 months of age
N response tto mycostatin,
No
t ti amphotericin,
h t i i
5-fluorocytosine
Addison’s disease & hypoparathyroidism
Anergic
g
Monocytopenic (including FCγ
FCγR+ cells)
High levels of IgG anticandida antibody
K F.
K.
F - pretreatment
p et eatment
Positive cultures for C. albicans
K F.
K.
F - pretreatment
p et eatment
- DCH to candida antigen
K F.
K.
F - pretreatment
p et eatment
Response to ttreatment
eatment
Peak monoccyte count (ce
ells/cumm)
Response to ttreatment
eatment
o
1500
1300
o
1100
900
700
P = <0.001
0 001
o
oo
o
0
2.5
5
7.5
TF dose (LEU x 109)
10
Response to T
Treatment
eatment
450
400
350
300
Control
Pre-Rx
Post-Rx
250
200
150
100
2.9 x 109 LEU TF
50
0
%
cumm
FCγR+ monocytes
%
Migration inhibition
K F.
K.
F – post
post--treatment
t eatment
+ DCH to candida antigen
K F.
K.
F – post
post--treatment
t eatment
Dend itic cells
Dendritic
Activate self
self--replicating (IL(IL-2 +) cytotoxic T
cells
Activate thymocytes
Activate T helper cells
Acti ate mac
Activate
macrophages
ophages
Promote TH1 differentiation
Kill viruses
i
(plasmacytoid
( l
t id DCs)
DC )
Detect PAMPs (TLRs)
Happier
Happie times
Congenital thymic disorders
DiGeorge syndrome
DiGeorge
DiGeo ge ssyndrome
nd ome
Immune deficiency due to single copy
deletion of TBX1
Encodes transcription factor TT-box
box--1 which
s required development of thymic
epithelium
DiGeorge syndrome
X
L H.
L.
H
23-day
23d -old
dayld baby
b b girl
il
Low calcium, parathyroid hormone
Absent thymic shadow
Truncus arteriosus Type IV
Characteristic phenotype
C didi i
Candidiasis
Imm ne assessment
Immune
Hypoergic
Marked T cell lymphopenia
Diminished mitogenic responses to PHA
Response to treatment
TF dose (LEU x 109)
3 646
3.646
4.80
5.49
2.56
10
5
0
10
20
30
40
50
60
Day of Treatment
70
80
0
0
Prolife
erative respo
onses (countss/min x 104
3.62
20
10
T Lymphocytes (%)
4.57
Control
Patient
Dend itic cells
Dendritic







Activate self
self--replicating (IL(IL-2 +) cytotoxic T
cells
Activate Thymocytes
Activate T helper cells
Acti ate mac
Activate
macrophages
ophages
Promote TH1 differentiation
Kill viruses
i
(plasmacytoid
( l
t id DCs)
DC )
Detect PAMPs (TLRs)
C
Congenital
i l CMV
C
L S.
L.
S
Positive urine & sputum cultures
L S.
L.
S
Imm ne assessment
Immune
Absent MIF production by CMVCMV-stimulated
PBMC
Patient
Control
Response to ttreatment
eatment
Post--treatment
Post
t eatment
MIF production in response to CMV
- CMV
+ CMV
Post--treatment
Post
t eatment
Negative urine & sputum cultures
Dend itic cells
Dendritic







Activate self
self--replicating (IL(IL-2 +) cytotoxic T
cells
Activate thymocytes
Activate T helper cells
Acti ate mac
Activate
macrophages
ophages
Promote TH1 differentiation
Kill viruses
i
Detect PAMPs (TLRs)
Happier
Happie times
Candida prosthetic valve
endocarditis
L. G.
Failed on two years of parenteral
antimicrobial therapy (amphotericin B,
B 55fluorocytosine)
R i f t d two
Reinfected
t
aortic
ti valve
l prostheses
th
Febrile, toxic, semicomatose, petechial
rash
Positive urine,, blood cultures for C. krusei
Imm ne assessment
Immune
Absent DCH to candida antigen
Leukopenia with 0% monocytes
Diminished MIF production with candida
antigen
High levels of IgG anticandida antibody
Response to treatment
25
20
wbc (per cumm)
15
monocytes (%)
10
2.48
2
48 x 109
LEU TF
migration
inhibition (%)
5
0
0
3
10
DAY OF TREATMENT
Blood cultures +
CH to candida 0
-
+
Response to treatment
+ C. krusei culture at 3 months
Dend itic cells
Dendritic







Activate self
self--replicating (IL(IL-2 +) cytotoxic T
cells
Activate thymocytes
Activate T helper cells
Acti ate mac
Activate
macrophages
ophages
Promote TH1 differentiation
Kill viruses
i
(plasmacytoid
( l
t id DCs)
DC )
Detect PAMPs (TLRs)
Chronic active hepatitis
B JJ.
B.
8 yearyear-old girl with biopsybiopsy-proven chronic
active hepatitis
p
B JJ.
B.
baseline
6 weeks
3 months
OT
330
850
320
PT
370
600
360
orexia
+
+++
+
0
+
0
0
+
0
er
dominal pain
Two closelyy spaced
p
doses of TF totaling
g 4.8 billion LIU given
g
at baseline
Disseminated molluscum
contagiosum
Atopic dermatitis
P. R.
Severe generalized eczema
Serum IgE >1,000 IU/mL
+ RAST to inhalants,, foods
Recalcitrant disseminated molluscum
P. R.
No response
esponse to high dose TF
Dend itic cells
Dendritic
Activate self
self--replicating (IL(IL-2 +) cytotoxic T
cells
 Activate thymocytes
 Activate T helper cells
 Activate
Acti ate mac
macrophages
ophages
 Promote TH1/Th2 differentiation
x Kill viruses
i
(plasmacytoid
( l
t id DCs)
DC )
 Detect PAMPs (TLRs)

Malignancies
Conclusions
Concl sions
TF th
therapy shows
h
promise
i as a treatment
t t
t
for congenital thymic disorders, and as an
adjuvant
dj
t iin th
the treatment
t t
t off certain
t i
recalcitrant viral and fungal infections in
selected
l t d patients
ti t
Further studies of its molecular contents,
modus operandus & clinical efficacy are
warranted
Sherwood
She ood Lawrence
La ence & TF
L
Low
molecular
l
l weight
i ht “informational”
“i f
ti
l” or
“activator” molecule
Gene dede-repressor?