Re: elevated cytokines

[Guest guest]

(and all others who might be interested),

I just found this article that I think you will find interesting (they even

mention steroid treatment in Section 12). It is called...

" FAQs: The meaning of neuroinflammatory findings in autism "

Here's the link...

http://www.neuro.jhmi.edu/neuroimmunopath/autism_faqs.htm

It's from s Hopkins but doesn't have any particular person's name on it.

I don't think it directly answers your question on cytokines, and frankly

it's so far over my head I don't even understand half of it, but I think you

might find it a good read.

Caroline

> From: " K. Fischer " <elfischer@...>

> Reply-< >

> Date: Thu, 30 Mar 2006 09:08:52 -0600

> < >

> Subject: RE: elevated cytokines

>

> So does this article then indicate that these children are in an

> autoimmune state or that they just have an unbalanced and chronically

> activated immune system? Will someone comment that knows the science

> better than I do. Thanks.

>

>

[Guest guest]

I was wondering if any similar findings were published

on Chronic Fatigue Syndrome patients?

--- Caroline Glover <sfglover@...> wrote:

> (and all others who might be interested),

>

> I just found this article that I think you will find

> interesting (they even

> mention steroid treatment in Section 12). It is

> called...

>

> " FAQs: The meaning of neuroinflammatory findings in

> autism "

>

> Here's the link...

>

>

http://www.neuro.jhmi.edu/neuroimmunopath/autism_faqs.htm

>

> It's from s Hopkins but doesn't have any

> particular person's name on it.

>

> I don't think it directly answers your question on

> cytokines, and frankly

> it's so far over my head I don't even understand

> half of it, but I think you

> might find it a good read.

>

> Caroline

>

>

> > From: " K. Fischer " <elfischer@...>

> > Reply-< >

> > Date: Thu, 30 Mar 2006 09:08:52 -0600

> > < >

> > Subject: RE: elevated cytokines

> >

> > So does this article then indicate that these

> children are in an

> > autoimmune state or that they just have an

> unbalanced and chronically

> > activated immune system? Will someone comment

> that knows the science

> > better than I do. Thanks.

> >

> >

>

>

>

__________________________________________________

[Guest guest]

this would be zimmerman/vargas/pardo's research item ...

doris

>

> (and all others who might be interested),

>

> I just found this article that I think you will find interesting

(they even

> mention steroid treatment in Section 12). It is called...

>

> " FAQs: The meaning of neuroinflammatory findings in autism "

>

> Here's the link...

>

> http://www.neuro.jhmi.edu/neuroimmunopath/autism_faqs.htm

>

> It's from s Hopkins but doesn't have any particular person's

name on it.

>

> I don't think it directly answers your question on cytokines, and

frankly

> it's so far over my head I don't even understand half of it, but I

think you

> might find it a good read.

>

> Caroline

>

>

> > From: " K. Fischer " <elfischer@...>

> > Reply-< >

> > Date: Thu, 30 Mar 2006 09:08:52 -0600

> > < >

> > Subject: RE: elevated cytokines

> >

> > So does this article then indicate that these children are in an

> > autoimmune state or that they just have an unbalanced and chronically

> > activated immune system? Will someone comment that knows the science

> > better than I do. Thanks.

> >

> >

>

[Guest guest]

>

> I was wondering if any similar findings were published

> on Chronic Fatigue Syndrome patients?

>

>Clinical & Experimental Immunology

Volume 142 Page 505 - December 2005

doi:10.1111/j.1365-2249.2005.02935.x

Volume 142 Issue 3

1: Clin Exp Immunol. 2005 Dec;142(3):505-11. Related Articles,

Links

Chronic fatigue syndrome is associated with diminished intracellular

perforin.

ORIGINAL ARTICLE

Chronic fatigue syndrome is associated with diminished intracellular

perforin

K. J. Maher*, N. G. Klimas* and M. A. Fletcher*

Summary

Chronic fatigue syndrome (CFS) is an illness characterized by

unexplained and prolonged fatigue that is often accompanied by

abnormalities of immune, endocrine and cognitive functions.

Diminished natural killer cell cytotoxicity (NKCC) is a frequently

reported finding. However, the molecular basis of this defect of in

vitro cytotoxicy has not been described. Perforin is a protein found

within intracellular granules of NK and cytotoxic T cells and is a

key factor in the lytic processes mediated by these cells.

Quantitative fluorescence flow cytometry was used to the

intracellular perforin content in CFS subjects and healthy controls.

A significant reduction in the NK cell associated perforin levels in

samples from CFS patients, compared to healthy controls, was

observed. There was also an indication of a reduced perforin level

within the cytotoxic T cells of CFS subjects, providing the first

evidence, to our knowledge, to suggest a T cell associated cytotoxic

deficit in CFS. Because perforin is important in immune surveillance

and homeostasis of the immune system, its deficiency may prove to be

an important factor in the pathogenesis of CFS and its analysis may

prove useful as a biomarker in the study of CFS.

Introduction Go to: ChooseTop of pageIntroduction <<Materials and

methodsResultsDiscussionConclusionsAcknowledgementsReferences

Chronic fatigue syndrome (CFS) is an illness characterized by

unexplained fatigue that persists for greater than 6 months, is not

alleviated by rest, and is associated with at least four of eight

case-defining symptoms such as impaired memory or concentration, sore

throat or lymph nodes, myalgia or arthralgia [1]. Other conditions

that could explain the illness must be excluded. The syndrome is

estimated to affect over 800 000 peoples in the United States,

although most are not diagnosed [2,3]. People with CFS are often sick

for many years [4]; the prognosis for recovery is poor [4,5]. The

lack of pathognomonic signs or accepted biomarkers contributes to the

difficult nature of the study and clinical management of CFS. A

concensus regarding the aetiology and pathophysiology of CFS has not

yet developed. However, the concept of immune dysfunction remains

actively debated, with reports of alterations in lymphocyte function,

activation and subset distributions common [6,7]. Frequently,

patients with chronic fatigue are found to have reduced cytotoxic

activity of natural killer (NK) cells (NKCC) in vitro[6–9]. Often,

the absolute count of NK cells is also low. However, we showed

reduced killing when effector cell concentration was defined as the

number of CD56+ CD3 lymphocytes in the assay and cytotoxic activity

was expressed as percentage killing of target cells at an effector to

target cell ratio of 1 : 1 [6,9]. Factors that contribute to reduced

NKCC have not been studied in the context of CFS. The purpose of the

present study was to investigate NK cell function and intracellular

concentration of the lytic granule protein perforin in NK cells in a

cohort of patients with CFS compared to matched sedentary but healthy

controls. The importance of perforin in cytotoxic function led to its

selection for study. Because of the decreased lytic activity of NK

cells from patients with CFS, we hypothesized that the intracellular

perforin content of NK cells would be reduced in CFS patients.

Materials and methods Go to: ChooseTop of pageIntroductionMaterials

and methods

<<ResultsDiscussionConclusionsAcknowledgementsReferences

Patients

The CFS study population consisted of 30 individuals (25 female, five

male; mean age years ± s.d. 46 ± 10) with symptoms that met the

Centers for Disease Control and Prevention (CDC)'s 1994 case

definition for CFS [1]. The mean age of onset was 36 ± 12 years.

Twenty-five subjects reported acute onset of the illness, while four

reported gradual onset. The mean duration of illness was 10 ± 7

years. The comparison group consisted of 19 apparently healthy but

sedentary (no regular exercise programme) controls (16 female, three

male; mean age years ± s.d. 43 ± 10). All subjects signed the

informed consent and the University of Miami's Internal Review Board

approved this protocol.

Cell surface phenotyping by flow cytometry

Ethylenediamine tetraacetic acid (EDTA) anti-coagulated whole blood

was surface-stained with optimal dilutions of CD45-FITC, CD14-PC5,

CD19-ECD, CD2-FITC, CD3-FITC, CD56-PE, CD4-ECD, CD8-PC5 and CD26-PE

and isotype controls in four colour combinations for 15 min at 25°C.

Samples were then fixed and lysed with Optilyse-C reagent, followed

by analysis on an XL-MCL flow cytometer. All reagents and

instrumentation were from the Beckman Coulter Corporation (Hialeah,

FL, USA). The accuracy and precision of analyses were optimized

through the adherence to the CDC's recommendations for flow

cytometric analyses [10].

Flow cytometric assessment of intracellular perforin

We have developed a flow cytometric method for the semiquantitative

assessment of intracellular perforin. The details of this method have

been published previously [11]. Heparinized whole blood (1 ml) was

fixed with 1 ml 4% p-formaldehyde (p-fma) for 15 min, followed by the

addition of 1 ml 17·5% bovine serum albumin (BSA), 0·1% NaN3 in

phosphate buffered saline (PBS) for 10 min to stop fixation. The

sample was washed twice with PBS containing 0·1% BSA, 0·1% NaN3.

Fixed samples were then resuspended in PBS containing 0·1% saponin

(Sigma, St Louis, MO, USA) for 10 min and washed twice in PBS with

0·1% saponin. Aliquots of fixed and permeabilized cells (50 µl) were

surface-stained for 15 min with optimal concentrations of CD8-FITC,

CD3-ECD and CD56-PC5 antibodies (Beckman Coulter). Isotypic control

or anti-perforin-phycoerythrin (PE) antibodies (Pharmingen, San

Diego, CA, USA) were then added to separate aliquots of the surface

stained cells for 30 min, followed by two saponin washes, one PBS

wash and one p-fma wash (0·1%). Samples were analysed on an XL-MCL

flow cytometer. Analyses consisted of a sequential gating strategy

whereby lymphocytes, defined by forward- and side-scatter properties,

were then phenotyped as CD3 CD56+ (NK) cells or CD3+ CD8+ (cytotoxic

T) cells. The anti-perforin-PE fluorescence intensity was measured in

the flow cytometer for each of these subsets and recorded as the

median of the fluorescence channel. Semiquantitative determinations

of cell-associated perforin were then derived from these median

fluorescence intensity values. Through the use of QuantiBRITE

fluorescence standards (Becton Dickinson), a standard curve was

calculated. These standards consist of a mixture of beads which are

impregnated with graded levels of PE and each bead set comes with

defined values (molecules PE/bead). Once these beads are analysed in

the flow cytometer, a defined number of molecules of PE can be

assigned to the median channel for the bead. From these channel

numbers and assigned values, a standard curve is generated and used

to convert median fluorescence intensity values for antiperforin-PE

binding to relative molecules perforin (rMol P)/cell.

NK cell cytotoxicity assay

The whole blood assay used relates the cytolytic activity of the

blood sample to the number of cells in the sample that are

phenotypically NK cells, as determined by flow cytometry and

lymphocyte count [12]. The assay was performed in triplicate at four

target to effector cell ratios. Heparinized blood was collected

before noon and transported to the laboratory at ambient temperature

so that the assay was started within 4 h of draw. An EDTA tube was

collected for the complete blood count that was needed for the

lymphocyte count. Target cells for the assay were the K562 cell line,

which was grown to log phase and then labelled with 51Cr. Labelled

target cells at final concentrations of 2, 1, 0·5 and 0·25 × 106

cells/ml were prepared and placed in triplicate wells of 96-well flat-

bottomed plates and 150 µl of whole blood were added. The spontaneous

release control was target cells plus 150 µl of assay medium and

total release control was target cells plus 150 µl of 1% Triton X-

100. The covered microtitre plate was centrifuged for 10 min at room

temperature at 400 g and incubated for 4 h at 37°C in 5% CO2

humidified atmosphere. Incubation was terminated by dispensing 100 µl

chilled (4°C) assay medium to each well. The plate was centrifuged

for 10 min at 400 g. Supernatant fluid (100 µl) was transferred to

counting tubes and counted in a gamma counter. To relate the

cytotoxic actvity of the blood sample to the cells in the sample that

were phenotypically NK cells, the lymphocyte count and four-colour

flow cytometry were performed to determine the number of CD56+ CD3

cells. Calculation of percentage cytotoxicity (CYT) for each dilution

of labelled target cells was performed as follows:

CYT = ((ER- & #8722; (SR-b)/(TR- & #8722; (SR-) × 100

where ER = mean counts per minute (cpm) of experimental release, SR =

mean cpm of spontaneous release, TR = mean cpm of total release and b

= gamma counter instrument backgound in cpm.

From the CYT of target cells killed and the number of target cells in

the well, with the number of effector cells (CD56+ CD3 lymphocytes)

held constant, a regression analysis was performed to calculate NKCC

(the percentage of cytotoxicity at an effector to target ratio of 1 :

1).

Statistics

All statistical comparisons were made using Student's t-test. Results

are presented as mean ± standard error of the mean (s.e.m.).

Correlations were made using the Pearson product–moment test.

Results Go to: ChooseTop of pageIntroductionMaterials and

methodsResults <<DiscussionConclusionsAcknowledgementsReferences

Standard surface flow cytometric phenotyping demonstrated that the

CFS group had a significantly reduced number of CD3 CD56+ (NK)

cells/µl of blood compared to the control group (122 ± 10 and 184 ±

27, respectively; P = 0·04). No differences were found between the

subject groups for the percentage of CD3 CD56+, CD3+ CD4+, CD3+ CD8+,

CD19+, CD3+ or CD2+ lymphocytes. The lymphocyte (T cells + NK cells)

compartment of the CFS group was found to be activated, as evidenced

by a significant elevation in the percentage of CD2+ CD26+ subset

when compared to healthy controls (58% ± 3 and 38% ± 3, respectively;

P = 0·001; Fig. 1). The number of CD2+ CD26+ lymphocytes/µl of blood

was also elevated in CFS patients (1069 ± 122 and 779 ± 66,

respectively; P = 0·05; Fig. 2). Consistent with the preponderance of

previous reports in the literature, the cytotoxic activity of natural

killer cells from subjects with CFS (21% ± 2 of K562 cells killed at

an effector to target cell ratio of 1 : 1) compared to healthy

controls (39% ± 5) was significantly lower (P = 0·001), as shown in

Fig. 3. In order to begin our study of NK function on the molecular

level, we used intracellular staining techniques to make quantitative

fluorescence measures of intracellular perforin. Perforin in NK cells

was found to be significantly reduced in CFS subjects when compared

to controls (3320 ± 313 and 6051 ± 956 rMol P/NK cell, respectively,

P = 0·01 (Fig. 4). Two representative analyses demonstrating the

difference in fluorescence intensity of antiperforin binding is shown

in Fig. 5 wherein the CFS subject had a lower level of anti-perforin

binding than the healthy control. Analysis of the perforin content of

the CD3+ CD8+ cytotoxic T cell subset revealed a reduction of

intracellular perforin content in the CFS group relative to controls

that approached statistical significance (270 ± 69 and 899 ± 309 rMol

P/Tc cell, respectively, P = 0·06 (Fig. 6). The correlation of rMol

P/NK cell and NKCC at an effector to target cell ratio of 1 : 1 for

all individuals studied (CFS subjects plus controls) was significant

(P = 0·01), as shown in Fig. 7.

Discussion Go to: ChooseTop of pageIntroductionMaterials and

methodsResultsDiscussion <<ConclusionsAcknowledgementsReferences

Many of the symptoms of CFS are inflammatory in nature (myalgia,

arthralgia, sore throat, tender lymphadenopathy), and have prompted a

theory of infection induced illness. CFS often presents with acute

onset of illness (reported in 60–80% of published samples) with

systemic symptoms similar to influenza infection that do not subside

[13]. However, reports of associated microbial infection or of latent

virus reactivation have been inconsistent. Whether associated with a

known antigen (e.g. a specific infection) or not, there is a

considerable literature describing immune activation in CFS. These

reports have described the elevation of lymphocyte surface activation

markers [6,14], the expression of proinflammatory cytokines and

evidence of Th2 cytokine increase [15–17]. In order to determine if

the cohort of CFS patients in this study had evidence of immune

activation we elected to analyse the surface marker, CD26 (dipeptidyl

peptidase IV). This ectoenzyme is known to increase upon cell

activation [18]. DPPIV/CD26 is a multifunctional molecule that is a

proteolytic enzyme, receptor, co-stimulatory protein, and is involved

in adhesion and apoptosis. CD26 is associated on T cells with

adenosine deaminase (ADA), and plays a major role in immune response.

Abnormal expression is found in autoimmune diseases, HIV-related

diseases and cancer [19]. Compared to controls, the CFS patients we

studied had a significantly elevated percentage and absolute count of

CD26+ lymphocytes.

The laboratory finding reported with the greatest consistency in CFS

patients is that of reduced NKCC [7,9]. In the few studies that

failed to find depressed NK activity in CFS subjects, methodological

problems may have been responsible [7]. In the present study,

consistent with previous reports, the NK-mediated cytotoxicity of CFS

subjects against the K562 cell targets was significantly lower than

that of controls. Although the CFS subjects in this cohort also had

significantly lower numbers of NK cells (CD56+ CD3) per volume of

blood, the diminished cytotoxicity was due to a decreased functional

capacity of the NK cells, as the percentage of killing in the 51Cr

release assay was calculated for both subject groups at a 1 : 1

target to CD56+ CD3 effector cell ratio. NK cells are critical for

immune surveillance against fungal, bacterial and viral infections.

They also play a vital role in cellular resistance to malignancy and

tumour metastasis [20]. NK cells can destroy their target cells by

calcium-dependent release of cytolytic granules, by activation of the

Fas (CD95) pathway or through contact with tumour necrosis factor

(TNF)- & #945;[21]. Perforin is released along with granzymes, particularly

granzyme B, from intracellular vesicles of cytolytic effector cells

and facilitates passage of these molecules through target cell

membranes, which then activate the apoptotic pathways of the caspases

[21]. NK cells differ from the other cytotoxic effector cell types

(CTL) in two major ways: they kill the target cells in a non-major

histocompatibility complex (MHC)-restricted fashion without the need

for previous in vitro or in vivo activation, and only NK cells

express constitutively the lytic machinery [20,21].

There was a recent suggestion in the literature of perforin reduction

in CFS. Steinhau et al. [22] used differential display polymerase

chain reaction (PCR) to search for candidate biomarkers in CFS. RNA

expression profiles of one subject with CFS and an age- and sex-

matched control showed differential expression of 10 genes. Of these,

five were down-regulated and one was perforin. In the present report,

we found that the relative number of molecules of perforin per NK

cells from CFS patients was significantly below that found in matched

healthy controls. This finding added support to the concept of an NK-

associated immune deficit in CFS and suggested that the decrease in

cytolytic potential of NK cells might be associated with a reduction

in the cell-associated concentrations of the effector molecule

perforin. A similar finding, which approached statistical

significance (P = 0·06), was a deficit in the perforin content of the

cytotoxic T subset. The observed decrease of perforin in T cells was

of an even greater magnitude than that seen in NK cells (30% and 55%

of control levels for Tc and NK, respectively). To our knowledge,

this is the first report of evidence to suggest a deficit in the

cytotoxic T cell compartment of CFS patients. These findings have

considerable significance in providing a potential mechanism in the

pathogenesis of CFS.

Two areas of research provided important insight in the role of

perforin in normal and pathological conditions. The perforin knockout

mouse, a model of perforin deficiency, demonstrated the role of

perforin as a cytotoxic effector molecule through decreased

cytotoxicity against virus infected and allogeneic targets [23] and

reduced clearance of intracellular pathogens and tumours [24,25].

These mice had increased numbers of activated CD8 cells [26] and

altered cytokine production with elevated expression of interleukin

(IL)-1, interferon (IFN)- & #947; and TNF- & #945;[27–29]. In the absence of

perforin, the granule protein granzyme A was proinflammatory, and

induced IL-6 and IL-8 expression [30].

A second area of investigation that pointed to the significance of

perforin in health and disease comes from studies of humans with the

condition known as familial haemophagic lymphohistiocytosis (FHL).

This rare and fatal disorder of early childhood was associated with a

genetic mutation that, in the homozygous state, resulted in the

absence of perforin expression [31]. Individuals with FHL were found

to suffer from an extensive immune activation, expansion and

infiltration of activated lymphocytes throughout the body, increased

expression of proinflammatory cytokines (IFN- & #947;, TNF- & #945;, IL-1 and IL-6)

and severely impaired cytotoxic abilities [32–34].

Given the deficiency of perforin among patients with CFS reported

here and the role of perforin in immune surveillance and

immunomodulation, we suggest that decreased intracellular perforin

content may play a role in the pathogenesis of CFS, an illness

reported to be associated with increased immune activation,

inflammatory cytokine levels, herpes virus reactivation and decreased

cytotoxicity. The mechanism responsible for the reduced perforin in

CFS remains unknown. The mechanisms may include genetic deficiency or

chronic microbial activation leading to perforin consumption and

exhaustion. An example of the latter mechanism is found in the report

of reduced NKCC and reduced intracellular perforin in patients with

chronic hepatitis C virus infection [35]. CFS is an illness whose

symptomatic expression is variable and any hypothosis regarding the

development of CFS should account for this variability. It can be

envisioned that either mechanism mentioned above could be subject to

variability. The diversity of symptoms may be due to the degree of

perforin deficiency, as a gene dosage effect is seen in mice [36] and

humans [34]; those who are heterozygous for the deficiency display an

intermediate phenotype. Secondly, exposure to activating stimuli may

be necessary for the evolution of symptomatic CFS in that perforin

deficiency in both children and C57BL/6 perforin knockout mice is not

associated with pathology until a viral infection stimulates an

uncontrolled immune activation [36,37]. Thus, superimposition of

immune activation, either microbial, autoimmune or even allergic, may

vary between those susceptible and alter the course of illness.

Thirdly, lymphocytes have multiple mechanisms to mediate killing. The

perforin pathway of cytotoxicity is distinct from that mediated by

Fas or TNF- & #945;. Perforin is essential for both lytic and granzyme-

mediated apoptotic killing but not Fas-mediated killing [21]. Because

these systems provide a similar function, varying levels of

redundancy among patients may explain varying degrees of

susceptibility to developing symptomatic illness.

Conclusions Go to: ChooseTop of pageIntroductionMaterials and

methodsResultsDiscussionConclusions <<AcknowledgementsReferences

We have presented evidence of a significant reduction in the

intracellular content of perforin among patients with chronic fatigue

syndrome. This molecule plays an important role in immune

surveillance against microbes and neoplasia as well as in immune

homeostasis. Such a deficiency is likely to be associated with

altered immune function and we would propose that this finding may be

important in the pathogenesis of CFS. However, this pathogenic

process is probably multifactorial, and variations in susceptibility

and inciting stimuli may account for the constellation of symptoms

seen in CFS. Perforin deficiency may prove useful as a biological

marker in CFS & #8722; perhaps one that will help define a subgroup with a

common pathogenesis.

Acknowledgements Go to: ChooseTop of pageIntroductionMaterials and

methodsResultsDiscussionConclusionsAcknowledgements <<References

This work was funded by support from the NIH Center Grant 1UD 1-AI

45940, the Miami Veterans Affairs Research and Education Foundation

and the CFIDS Association of America.

References Go to: ChooseTop of pageIntroductionMaterials and

methodsResultsDiscussionConclusionsAcknowledgementsReferences <<

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Clinical & Experimental Immunology

Volume 142 Page 505 - December 2005

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Authors:

K. J. Maher

N. G. Klimas

M. A. Fletcher

Keywords:

chronic fatigue syndrome

cytoplasmic granules

flow cytometry

killer cells – natural

perforin

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Accepted for publication 8 August 2005

Affiliations

*Department of Medicine, University of Miami School of

Medicine, Miami, FL, USA, and Department of Medicine, Veterans

Administration Medical Center, Miami, FL, USA

Correspondence

Dr Ann Fletcher, Department of Medicine R-42, University of

Miami School of Medicine, 1600 NW 10th Avenue, Miami, FL

33176, USA.

E-mail: mfletche@...

Image Previews

[Full Size]

Fig. 1. Flow cytometry indicted that the percentage of lymphocytes [T

cells and natural killer (NK) ce...

[Full Size]

Fig. 2. Flow cytometry indicated that the number of CD2+ lymphocytes

[T cells and natural killer (NK) ...

[Full Size]

Fig. 3. Subjects with chronic fatigue syndrome had significantly

lower natural killer (NK) cytotoxicit...

[Full Size]

Fig. 4. Subjects with chronic fatigue syndrome had significantly

lower intracellular perforin in natur...

[Full Size]

Fig. 5. Representative histograms showing the fluorescence intensity

of the antiperforin antibody bind...

[Full Size]

Fig. 6. Subjects with chronic fatigue syndrome had a lower

intracellular cytotoxic T cell perforin lev...

[Full Size]

Fig. 7. There was a significant correlation between natural killer

(NK) cell activity (NKCC) and amoun...

To cite this article

Maher, K. J., Klimas, N. G. & Fletcher, M. A. (2005)

Chronic fatigue syndrome is associated with diminished intracellular

perforin.

Clinical & Experimental Immunology 142 (3), 505-511.

doi: 10.1111/

j.1365-2249.2005.02935.x

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[Guest guest]

>

> I was wondering if any similar findings were published

> on Chronic Fatigue Syndrome patients?

>

Dr. Goldberg & Mena's early work found similar findings on Spect as

well.

Elyse Goldberg

> NEUROSPECT: ASSESSMENT OF ABNORMAL DISTRIBUTION OF RCBF IN CFIDS

VS. " AUTISTIC SYNDROME CHILDREN. "

Goldberg, Ismael Mena, Bruce. , and Carmen .

Dept. of Nuclear Medicine, Imaging Center, Harbor UCLA Medical

Center, Torrance, Ca.

OBJECTIVE: To compare NeuroSpect finding in 25 children

diagnosed " Autistic Syndrome / PDD " with 13 children with CFS / CFIDS

vs. normals.

METHODS: We report on quantitative (Xe133) rCBF and high resolution

HMPAO SPECT in 25 children meeting criteria of DSM IV for autism,

compared with 13 children meeting CDC criteria for chronic fatigue

syndrome (CFIDS) and 13 normal children (HMPAO). Ages were 5.5+2.5

years, 13+3 years and 9.3+3.2 years respectively. Male/female ratios

were 22/3, 7/6 and 8/5 respectively. RCBF was imaged with a brain

dedicated imaging device (Shimatzu, Headtome) after inhalation of

1.110 MBq of Xe133 gas, and with a high resolution fan beam

collimator after IV injection of 370 - 740 MBq of Tc99m HMPAO. ROI's

were determined manually for Xe133 and automatically set for HMPAO

(64/ transaxial cut, in 6 adjacent 1 cm cuts above basal ganglia).

RESULTS:

Xe133 rCBF (ml/min/100g)

Max. Flow Min. Flow Avg. Flow

1. Autism 116+28 ** 49+10 * 92+22 *** lvs2

2. CFIDS 86+11 35+5 63+7

**lvs3

3. Normals 62+9x

p<0.001 *** x Chiron et al., J.Nuc. Med;

1992:33,696-703

0.002 **

0.02 *

In the Autistic children, maximal rCBF was observed in frontal lobes,

while minimal rCBF was detected in temporal and occipital lobes and

cerebellum. HMPAO uptake was 0.50+5 in occipital lobes and in

frontal lobes 0.82+4, p<0.0001, while in Normals it was 0.78+5,

without significant gradient. In the CFIDS children, hypoperfusion

is observed at 42 + 10 ml/min/100g, p < 0.0001 in the left temporal

lobe and at 45 + 11, p < .001 in right temporal lobe. There is

furthermore hypoperfusion with similar statistical significance in

both parietal lobes and at 50 and 53 ml/min/100g, p < 0.05 in the

frontal lobe of the right hemisphere.

SUMMARY: Brain Spect Scan results are presented along with some

clinical observations of these particular groups of patients. This

tool may open the door to a more physiologic/medical approach to this

process in children. Comparisons are made with the finding in

children with CFIDS and those " labeled " Autistic Syndrome / PDD. The

observation of temporal hypoperfusion in adults and children with

CFS/CFIDS, may help define Autism as a disorder of impaired relations

with the surrounding environment determined by the temporal

hypofunction leading as a consequence to a diaschetic hypofunction of

visual cortex and cerebellum. The mechanisms for this abnormality

need to be investigated using activation techniques and other

approaches i.e.: evaluation of possible immune dysregulation, etc.