chronic fatigue syndrome, cfs, chronic fatigue immune dysfunction, cfids, myalgic encephalopathy, fibromyalgia, fms, Forum,  myalgic encephal, m.e., low blood volume, neurotoxin, orthostatic intolerance, multiple chemical sensitiv, National CFIDS Foundation, chronic fatigue, NCF Forum newsletter, chronic illness

Part Two - Total Exposure: Expanded Model for RNase L Fragmentation in CFS Uncovered; The National CFIDS Foundation Announces the Use of Elastase Inhibitors as a Potential Treatment for CFS Patients

Investigative Report by Alan Cocchetto, Medical Director National CFIDS Foundation, Inc.
Copyright 2003 - Written Permission Required for Reprinting or Distributing

From our investigation, the NCF had also identified several additional patents that yielded new information that hadn't been seen by other scientists or physicians in the field. In one patent [23], Dr. DeMeirleir and his colleagues expanded the RNase L fragmentation model in CFS and provided descriptions for their scientific discoveries: "The lower molecular weight fragments of RNase L are produced from the specific cleavage of native RNase L by a protease(s). These fragments then play a selective role in inhibiting the apoptotic pathway, in effect keeping the damaged cell alive and the immune system dysfunctional." The researchers had identified three RNase L fragments. In Fragment #1, they found an ankyrin binding repeat domain which is known to interact with various transport proteins. In addition, they also identified sequence homology with
NF-kappaB (Nuclear transcription Factor kappa B).

NF-kappaB has been demonstrated to induce transcriptional activities within a cell to promote cell growth thus acting in an anti-apoptotic manner. In Fragment #2, they identified the 2-5A binding fragment that has catalytic activity and thus is able to degrade RNA. Since this fragment competes with the native RNase L protein for free 2-5A, it may be responsible for inhibiting the complete induction of apoptosis. They found that Fragment #3 shared homology with chain A of Cdk6 (Cyclin dependent kinase). Cdk6 acts to block apoptosis by altering the cell cycle. In fact, the inventors provide a diagram (Figure 1) that explicitly shows the role of these RNase L fragments in CFS pathogenesis. Since the 2-5A synthetase / RNase L pathway is part of the antiviral defense mechanism in cells and because it plays a role in the regulation of cell growth / differentiation, these RNase L fragments provide critically important insight into the cellular mechanisms that are uniquely responsible for the immune dysfunction in CFS patients.

Furthermore, the inventors conclude that "The presence (or absence) of low molecular weight RNase L fragments 1 to 3, and optionally the caspase activity data, is then used to diagnose whether or not the host suffers from the chronic immune disease." Caspases are cysteine-aspartic acid proteases that are considered to be the cell death executioners due to the absolute requirements for their presence in cell death / apoptosis [24].

Extensive analysis was done by Dr. DeMeirleir and his colleagues in this patent. They studied four patients who had been administered ampligen for a period of at least six months. These patients were also compared with healthy controls and forty-four additional patients who had not undergone ampligen therapy. All patients were evaluated for RNase L fragmentation and caspase levels. For the 44 patients, when the RNase L ratios were 6 to 8 (6 < RNase L ratio < 8), the caspases C2, C3, C6, C8, and C9 reached their peak. Caspase levels were generally lower overall for lower RNase L levels (RNase L ratio < 6) as well as for higher RNase L levels (RNase L ratio > 8). In fact the inventors concluded "Upon further analysis, it is evident that levels of all caspases assayed (C3, C3, C6, C8, and C9) at first increase, then decrease relative to the RNase L ratio in CFS patient groups P1 through P7, indicating that the apoptotic process is being inhibited when the levels of RNase L-related fragments reaches a certain point." In their evaluation of four patients treated with ampligen, two patients responded to therapy in that their RNase L ratio decreased and they experienced increases in their Karnofsky performance scores (KPS). The Karnofsky performance test measures an individual's ability to function and carry on normal activities. Karnofsky scores range from zero for a nonfunctional or dead patient to 100 for a completely normal person. The third patient did not respond to ampligen due to an increase in the RNase L ratio and a decrease in Karnofsky score. The last patient responded to ampligen initially with reductions in RNase L ratio and an increase in Karnofsky score. However, six months after cessation of therapy, the patient suffered a relapse as indicated by a decrease in Karnofsky score and a significant increase in the RNase L ratio. Interestingly, for this patient, the RNase L ratio after the cessation of therapy and subsequent relapse was much greater (52.9) than even before beginning therapy (8.7). In both the non-responder and relapsed patient, low levels of caspase-3 (C3) that were found may indicate that a block exists in the apoptotic pathway.

Dr. DeMeirleir and colleagues, in this patent, concluded that (A) Increases or decreases in the relative amounts (i.e., ratios) of native RNase L when measured against RNase L-related fragments correlates strongly with the presence or absence of CFS disease, respectively; (B) Increases in apoptosis in PBMCs from CFS patients can be measured by analyzing caspase levels; (C) Increases in apoptosis in PBMCs from CFS patients can be measured by analyzing the relative amount of native RNase L and related fragments; (D) As the ratio of RNase L-related fragments to the remaining native RNase L protein increases above a certain level, the process of apoptosis, as measured by caspase levels, appears to stop then decline even further to sub-normal levels; (E) RNase L-related fragments are likely to inhibit the apoptotic process based on the amino acid sequence comparison of RNase L-related fragments to known inducers of cell activation and growth; (F) Upon successful therapy with mismatched double stranded RNA (i.e. ampligen), RNase L ratios return to 'normal' or healthy control levels; (G) Upon successful therapy with mismatched double stranded RNA, caspase levels return to 'normal' or healthy control levels; (H) Mismatched double-stranded RNA induces the synthesis of 2'-5'A. In turn, 2'-5'A binds to and activates native RNase L homodimers that in turn induce apoptosis, removing the anti-apoptotic block, allowing for return of normal cellular functions.

However, Dr. DeMeirleir and colleagues have provided exciting information regarding a potential treatment therapy identified in their newest patent. The patent, titled "Methods of treatment of chronic immune diseases," was just issued on July 31, 2003 [25]. In this patent, the inventors detailed their findings regarding treatment for multiple sclerosis and chronic fatigue syndrome, both of which the inventors consider to be chronic immune diseases. Treatment includes administering to a human an effective amount of a protease inhibitor where this protease inhibitor is an elastase inhibitor and where the elastase inhibitor is a beta-lactam containing compound such as the cephem cefoperazone. Furthermore, the inventors state that the diagnosis of chronic immune disease is accomplished by detecting the presence of RNase L fragments. This comes as no surprise because of the volumes of previous work on RNase L as well as the inventors previous patents. In fact, another research group has recently published that RNase L ratios could distinguish CFS patients from healthy controls [26].

Briefly stated, Dr. DeMeirleir and colleagues demonstrated that the enzyme elastase is able to generate fragments of recombinant RNase L protein, the size of which approximates the fragment of native RNase L protein found in peripheral blood mononuclear cells from patients with MS and CFS. This is in-line with a previous publication by DeMeirleir and others [27] where RNase L proteolysis could be mimicked by combining recombinant RNase L with human leukocyte elastase. In the patent, the inventors were able to demonstrate that pure / recombinant RNase L (> 95%) was proteolytically cleaved by elastase and the fragments of RNase L generated by elastase digestion were compared to the size of the fragments of native RNase L found in PBMCs from CFS patients. The inventors demonstrated that the pure RNase L cleavage product (37 kDa) was equivalent in size to that found in CFS patients. The inventors then tested the effects of the beta-lactam based antibiotic cefoperazone (Pfizer Pharmaceuticals, trade name Cefobid) on the levels of RNase L protein in the human monocytic leukemia cell line U937. The inventors demonstrate that cefoperazone is able to inhibit the production of the low-molecular weight fragment of native RNase L protein and that this inhibition is dose dependent. They also demonstrate that cefoperazone is able to inhibit elastase activity as indicated by a reduction of low-molecular weight RNase L protein by in-vitro testing. This indicates that beta-lactam based antibiotics have an effect on elastase, via its inhibition, and thereby are able to reduce fragmentation associated with RNase L cleavage. In fact, from the previous publication, the authors state that enhanced human leukocyte elastase activity appears to be involved in the increased proteolysis of RNase L in CFS PBMCs. Since the inventors listed numerous antibiotics that contain beta-lactam compounds, without discussing their targeted sensitivity, this could help explain why some patients are at least partially responsive to certain antibiotics and why they have reported that they feel better on antibiotic therapy. If their antibiotics contain beta-lactam based compounds then elastase would be inhibited and this would potentially move the patient in the direction of native RNase L restoration by reducting the cleavage products!

A brief look by the NCF into the role(s) of human leukocyte elastase provided us with answers to many additional questions that we had. One of these was the fact that elastase has been found in significant amounts in several different types of amyloidoses [28] and in amyloid fibrils themselves [29,30]. The NCF had noted previous research that examined the molecular basis of CFS [31]. In this work, a unique urinary marker contained N-methylpyrrolidine and was found to be highly significant in CFS patients. However, N-methylpyrrolidine has already been found to accelerate the aggregation process associated with amyloid formation [32]. These findings strengthen our NCF hypothesis for potential amyloid involvement and protein misfolding in the pathogenesis of CFS.

Elastase has been found to cleave all six isulin-like growth factor binding proteins (IGFBPs) in-vitro and in-vivo with a significant proteolytic cleavage of IGFBP-3 [33]. IGFBP's are associated with the proliferative effects of insulin-like growth factors (IGFs) on various cells. Interestingly, IGFBP-3 has been found to upregulate STAT1 and to increase its protein

expression [34]. Acclydine therapy, as suggested by Dr. DeMeirleir, acts by increasing IGFBP-3 [19]. Thus, by increasing the levels of IGFBP-3 in the cell, IGF-1 is blocked from binding to its receptor thereby suppressing the growth of the cell, promoting apoptosis, and counteracting the loss of functional p53 protein on the growth of the cell. Additionally, lycopene, which is a natural carotenoid found in tomatoes and readily available as a supplement, has also been found to increase IGFBP-3 as well [35,36].

Elastase has been implicated in chronic inflammation [37] including rheumatoid arthritis [38] and is the target of antirheumatic drugs [39]. In fact, structural changes have been observed in the skin [40] by elastase and in the kidney [41] elastase mediates glomerular injury in-vivo causing proteinuria due to changes in glomerular permeability. Elastase has also been found to cleave the T4-binding globulin (TBG) [42]. TBG serves to maintain an important serum pool of thyroid hormones and to prevent their excessive loss in urine. Elastase also cleaves the corticosteroid-binding globulin reducing its hormone-binding affinity and capacity [42]. In addition, elastase is involved in fibrinolysis [43] as it has been found to degrade fibrin and inhibit the blood coagulation system by degrading key proteins.

However, other interesting findings for elastase are that it regulates Stromal cell-derived factor-1 (SDF-1)/CXCR4 binding [44]. This is particularly intriguing because Human Herpes Virus-6 (HHV-6) uses the CXCR4 receptor for infection [45]. HHV-6 has been implicated in the pathology for both multiple sclerosis and chronic fatigue syndrome [46]. Furthermore, elastase has been found to cleave CD4 (helper cells) and CD8 (cytotoxic/suppressor cells) lymphocytes leading to a reduction in number of these cell types [47]. In addition, elastase has also been found to cleave immune complexes and to regulate inflammation by a feedback mechanism that can lead to cyclic inflammatory states [48].

The finding that Dr. DeMeirleir and colleagues found beta-lactam antibiotics to be associated with the inhibition of human leukocyte elastase is echoed by various publications [49]. However, the NCF has identified one sensitive inhibitor of leukocyte elastase that is readily available and it is boswellic acid [50].

Dr. DeMeirleir's colleagues also have a recent patent application for the cleavage of actin [51]. In this application, the inventors found correlations between the relative amount of native RNase L protein in PBMC extracts and the relative amount of native actin protein in serum. They also found correlations between the ratio of RNase L fragments and the ratio of actin fragments. Actin is a protein that assists in the diverse activities of the cytoskeleton of the cell which includes cell signalling. Interestingly, elastase has been shown to cleave actin in Behcet's disease, a disorder that involves inflammation of the blood vessels [52].

In conclusion, it is obvious that Dr. DeMeirleir and his colleagues, in cooperation with other medical researchers, have investigated several key mechanisms in the pathogenesis of CFS and MS and these included STAT1, RNase L cleavage, actin, and the use of elastase inhibitors as a potential therapy for these chronic diseases. Hopefully by highlighting this research and explaining how it fits within a larger disease framework, the NCF has acted in a responsible

manner to inform, educate, and to provide much needed hope to patients, their physicians, and to researchers worldwide.

Part two article review at a glance:

Dr. DeMeirleir and colleagues scientific finding:

  • Expanded model for RNase L fragmentation identified (three fragments) in CFS patients
  • Fragment #1 is identified with NF-kappaB mimicry
  • Fragment #2 is identified with 2'-5'A binding
  • Fragment #3 is identified with Cdk6 chain A mimicry
  • Apoptosis (cell death) increases initially in CFS and then is inhibited when the levels of RNase L related fragments reach a certain point.

Scientific interpretation:

  • Expanded RNase L model helps to explain immune system dysfunction in CFS.
  • Use of leukocyte elastase inhibitors may potentially treat patients with CFS as well as MS due to its impact on RNase L and STAT1.
  • Beta-lactam based antibiotics act as leukocyte elastase inhibitors, however drug sensitivity is most likely unknown for this target.

The National CFIDS Foundation, Inc. provides informative and up-to-date quality scientific research assessments for the CFS patient community, treating physicians, and medical researchers alike. By utilizing evidence-based medical techniques which incorporate the judicious use of intellectual property rights in conjunction with traditional disease profiling methods, investigational reports are generated with the explicit intent of validating the clinical importance and applicability of a discovery as well as to assist in propelling appropriate CFS medical research forward.

Additional details on all of our research activity and scientific results will be discussed in future issues of The Forum and updated on our website at We greatly appreciate all donors and supporters of the NCF's Research Grant Program. Without your financial support, we would be unable to appropriately fund the aggressive research mentioned in this article. However, since this research is ongoing, please consider continued donations to the NCF's Research Grant Program to support these efforts. The National CFIDS Foundation, Inc. is the largest, all volunteer, and fastest growing non-profit CFIDS organization in the U.S. Because we have no paid staff, 100% of all donations go directly to research. Please continue to help us help you by supporting us financially!

1. Chronic Fatigue Syndrome: A Biological Approach; Englebienne P, DeMeirleir K; CRC Press, 2002

2. The 2-5A Pathway and Signal Transduction: A Possible Link to Immune Dysregulation and Fatigue; Englebienne P, Herst CV, Fremont M, Verbinnen T, Verhas M, DeMeirleir K; 5: 99-130; in Chronic Fatigue Syndrome: A Biological Approach

3. STATs: Transcriptional control and biological impact; Levy DE, Darnell JE; Nature Reviews: Mol Cell Bio 2002; 3: 651-662

4. Impaired response to interferon-alpha/beta and lethal viral disease in human STAT1 deficiency; Dupuis S, Jouanguy E, Al-Hajjar S, Fieschi C, Al-Mohsen IZ, Al-Jumaah S, Yang K, Chapgier A, Eidenschenk C, Eid P, Al Ghonaium A, Tufenkeji H, Frayha H, Al-Gazlan S, Al-Rayes H, Schreiber RD, Gresser I, Casanova JL; Nat Genet 2003; 33(3): 388-391

5. STAT1 deficiency unexpectedly and markedly exacerbates the pathophysiological actions of IFN-alpha in the central nervous system; Wang J, Schreiber RD, Campbell IL; Proc Natl Acad Sci 2002; 99(25): 16209-16214

6. Targeted disruption of the mouse STAT1 gene results in compromised innate immunity to viral disease; Durbin JE, Hackenmiller R, Simon MC, Levy DE; Cell 1996; 84(3): 443-450

7. Targeted disruption of the STAT1 gene in mice reveals unexpected physiologic specificity in the JAK-STAT signalling pathway; Meraz MA, White JM, Sheehan KC, Bach EA, Rodig SJ, Dighe AS, Kaplan DH, Riley JK, Greenlund AC, Campbell D, Carver-Moore K, DuBois RN, Clark R, Aguet M, Schreiber RD; Cell 1996; 84(3): 431-442

8. The antitumor effects of IFN-alpha are abrogated in a STAT1-deficient mouse; Lesinski GB, Anghelina M, Zimmerer J, Bakalakos T, Badgwell B, Parihar R, Hu Y, Becknell B, Abood G, Chaudhury AR, Magro C, Durbin J, Carson III WE; J Clin Invest 2003; 112(2): 170-180

9. US Patent # 6,080,554; issued June 27, 2000; filed April 27, 1999; titled Methods and compositions for use in characterizing multiple sclerosis disease activity in a subject; Inventors: Campine IWL, DeMeirleir KL, Herst CVT; Assignee: R.E.D. Laboratories, NV.

10. US Patent # 6,214,554; issued April 10, 2001; filed April 21, 1999; titled Chronic fatigue syndrome diagnosis; Inventor: Suhadolnik RJ; Assignee: Temple University of the Commonwealth System of Higher Education

11. Low expression of interferon-stimulated genes in active multiple sclerosis is linked to subnormal phosphorylation of STAT1; Feng X, Petraglia AL, Chen M, Byskosh PV, Boos MD, Reder AT; J Neuroimmunol 2002 Aug; 129(1-2): 205-215

12. US Patent Application # 20030017493; published January 23, 2003; filed June 17, 2002; titled Methods for the detection and treatment of chronic immune diseases; Inventors: Fremont M, Englebienne P, Herst CVT

13. US Patent Application # 20030077674; published April 24, 2003; filed June 17, 2002; titled Methods for diagnosis and treatment of chronic immune diseases; Inventors: Fremont M, Englebienne P, Herst CVT

14. R.E.D. Laboratories website; ; website last updated: 1/16/01

15. Flemish Parliament of Belgium; dated June 6, 2001; Flemish Parliament Social Policy Note: Environment and Health, Hearings and Advice, Session 2000-2001, Article 740 (2000-2001); Testimony of Professor Kenny DeMeirleir, Human Physiology, VUB hearing of March 5, 2001

16. Prevention of beta-amyloid neurotoxicity by blockade of the ubiquitin-proteasome proteolytic pathway; Favit A, Grimaldi M, Alkon DL; J Neurochem 2000; 75(3): 1258- 1263.

17. The protease inhibitor, MG132, blocks maturation of the amyloid precursor protein swedish mutant preventing cleavage by beta-secretase; Steinhilb ML, Turner RS, Gaut JR; J Bio Chem 2001 Feb 9; 276(6): 4476-4484

18. Utility of the blood for gene expression profiling and biomarker discovery in chronic fatigue syndrome; Vernon SD, Unger ER, Dimulescu IM, Rajeevan M, Reeves WC; Dis Markers 2002 18:193-199

19. US Patent Application # 20030017492; published January 23, 2003; filed June 17, 2002; titled Methods for diagnosis and treatment of chronic immune diseases; Inventors: Fremont M, Englebienne P, Herst CVT

20. Immune Cell Apoptosis and Chronic Fatigue Syndrome; Fremont M, D'Haese A, Roelens S, DeSmet K, Herst CV, Englebienne P; 6: 131-174; in Chronic Fatigue Syndrome: A Biological Approach

21. The STAT family in cytokine signaling; Ihle JN; Current Opinion in Cell Bio 2001; 13: 211-217

22. The antitumor effects of IFN-alpha are abrogated in a STAT1-deficient mouse; Lesinski GB, Anghelina M, Zimmerer J, Bakalakos T, Badgwell B, Parihar R, Hu Y, Becknell B, Abood G, Chaudhury AR, Magro C, Durbin J, Carson III WE; J Clin Invest 2003 July, 112(2): 170-180

23. World Patent # WO0215929; issued February 28, 2002; filed August 16, 2001; titled Methods and compositions for use in the diagnosis and treatment of chronic immune disease; Inventors: Englebienne P, DeMeirleir KL, Herst CVT; Applicant: R.E.D. Laboratories, N.V.

24. Cell death program; Debatin K; in Textbook of Malignant Haematology; Degos L, Linch DC, Lowenberg B, eds. London UK: Martin Dunitz Ltd; 1999: 153-164

25. World Patent # WO03061605; issued July 31, 2003; filed January 10, 2003; titled Methods of treatment of chronic immune disease; Inventors: El Bakkouri K, Englebienne P, DeMeirleir K, Herst CVT; Applicant: R.E.D. Laboratories, N.V.

26. RNase L levels in peripheral blood mononuclear cells: 37-Kilodalton/83-Kilodalton isoform ratio is a potential test for chronic fatigue syndrome; Tiev KP, Demettre E, Ercolano P, Bastide L, Lebleu B, Cabane J; Clin Diag Lab Imm 2003; 10(2): 315-316.

27. Ribonuclease L proteolysis in peripheral blood mononuclear cells of chronic fatigue syndrome patients; Demettre E, Bastide L, D'Haese A, DeSmet K, DeMeirleir K, Tiev KP, Englebienne P, Lebleu B; J Biol Chem 2002; 277(38): 35746-35751.

28. Neutrophil proteases associated with amyloid fibrils; Stone PJ, Campistol JM, Abraham CR, Rodgers O, Shirahama T, Skinner M; Biochem Biophys Res Commun 1993; 197(1): 130-136

29. The association of an elastase with amyloid fibrils; Skinner M, Stone P, Shirahama T, Connors LH, Calore J, Cohen AS; Proc Soc Exp Biol Med 1986; 181(2): 211-214

30. Elastase-type proteases on the surface of human blood monocytes: possible role in amyloid formation; Lavie G, Zucker-Franklin D, Franklin EC, J Immunol 1980; 125(1): 175-180

31. Preliminary determination of a molecular basis of chronic fatigue syndrome; McGregor NR, Dunstan RH, Zerbes M, Butt HL, Roberts TK, Klineberg JJ; Biochem Mol Med 1996; 57(2): 73-80

32. Nicotine inhibits amyloid formation by the beta-peptide; Salomon AR, Marcinowski KJ, Friedland RP, Zagorski MG; Biochemistry 1996; 35(42): 13568-13578

33. Inflammation-related neutral proteases, cathepsin G and elastase, function as insulin- like growth factor binding protein proteases; Gibson TL, Cohen P; Growth Horm IGF Res 1999; 9(4): 241-253

34. Identification of STAT-1 as a molecular target of IGFBP-3 in the process of chondrogenesis; Spagnoli A, Torello M, Nagalla SR, Horton WA, Pattee P, Hwa V, Chiarelli F, Roberts DT Jr, Rosenfeld RG; J Biol Chem 2002; 277(21): 18860-18867

35. Lycopene supplementation inhibits lung squamous metaplasia and induces apoptosis via up-regulating insulin-like growth factor-binding protein 3 in cigarette smoke-exposed ferrets; Liu C, Lian F, Smith DE, Russell RM, Wang XD; Cancer Res 2003; 63(12): 3138-3144

36. Are dietary influences on the risk of prostate cancer mediated through the insulin-like growth factor system?; Mucci LA, Tamimi R, Lagiou P, Trichopoulou A, Benetou V, Spanos E, Trichopoulos D; BJU Int 2001; 87(9): 814-820

37. The role of neutrophil elastase in chronic inflammation; Doring G; Am J Respir Crit Care Med 1994; 150(6 Pt 2): S114-S117

38. Cathepsin G and elastase in synovial fluid and peripheral blood in reactive and rheumatoid arthritis; Nordstrom D, Lindy O, Konttinen YT, Lauhio A, Sorsa T, Friman C, Pettersson T, Santavirta S; Clin Rheumatol 1996; 15(1): 35-41

39. The inhibitory effects of antirheumatic drugs on the activity of human leukocyte elastase and cathepsin G; Steinmeyer J, Kalbhen DA; Inflamm Res 1996; 45(7): 324-329

40. Structural changes of human epidermis induced by human leukocyte-derived proteases; Ludolph-Hauser D, Schubert C, Wiedow O; Exp Dermatol 1999; 8(1): 46-52

41. The human neutrophil serine proteinases, elastase and cathepsin G, can mediate glomerular injury in vivo; Johnson RJ, Couser WG, Alpers CE, Vissers M, Schulze M, Klebanoff SJ; J Exp Med 1988; 168(3): 1169-1174

42. Characterization of T(4)-binding globulin cleaved by human leukocyte elastase; Janssen OE, Golcher HM, Grasberger H, Saller B, Mann K, Refetoff S; J Clin Endocrinol Metab 2002; 87(3): 1217-1222

43. The elastase-mediated pathway of fibrinolysis; Machovich R, Owen WG; Blood Coagul Fibrinolysis 1990; 1(1): 79-90

44. Leukocyte elastase negatively regulates Stromal cell-derived factor-1 (SDF-1)/CXCR4 binding and functions by amino-terminal processing of SDF-1 and CXCR4; Valenzuela- Fernandez A, Planchenault T, Baleux F, Staropoli I, Le-Barillec K, Leduc D, Delaunay T, Lazarini F, Virelizier JL, Chignard M, Pidard D, Arenzana-Seisdedos F; J Biol Chem 2002; 277(18): 15677-15689

45. Transcriptional down-regulation of CXC chemokine receptor 4 induced by impaired association of transcription regulator YY1 with c-Myc in human herpesvirus 6 infected cells; Hasegawa A, Yasukawa M, Saksi I, Fujita S; J Immunol 2001; 166(2): 1125-1131

46. Frequent HHV-6 reactivation in multiple sclerosis (MS) and chronic fatigue syndrome (CFS) patients; Ablashi DV, Eastman HB, Owen CB, Roman MM, Friedman J, Zabriskie JB, Peterson DL, Pearson GR, Whitman JE; J Clin Virol 2000; 16(3): 179-191

47. Cleavage of lymphocyte surface antigens CD2, CD4, and CD8 by polymorphonuclear leukocyte elastase and cathepsin G in patients with cystic fibrosis; Doring G, Frank F, Boudier C, Herbert S, Fleischer B, Bellon G; J Immunol 1995; 154(9): 4842-4850

48. Elastase from polymorphonuclear leukocytes: a regulatory enzyme in immune complex disease; Doring G, Goldstein W, Botzenhart K, Kharazmi A, Schiotz PO, Hoiby N, Dasgupta M; Clin Exp Immunol 1986; 64(3): 597-605

49. Mechanism of inhibition of human leucocyte elastase by monocyclic beta-lactams; Chabin R, Green BG, Gale P, Maycock AL, Weston H, Dorn CP, Finke PE, Hagmann WK, Hale JJ, MacCoss M et. al.; BioChemistry 1993; 32(34): 8970-8980

50. Inhibition by boswellic acids of human leukocyte elastase; Safayhi H, Rall B, Sailer ER, Ammon HP; J Pharmacol Exp Ther 1997; 281(1): 460-463

51. US Patent Application # 20030152919; published August 14, 2003; filed May 15, 2000; titled Method and compositions for use in diagnosing and characterizing chronic immune disease; Inventors: Roelens SAM, Englebienne P, D'Haese AMYR, Herst CVT

52. Characterization of a protease responsible for truncated actin increase in neutrophils of patients with Behcet's disease; Yamashita S, Suzuki A, Yanagita T, Hirohata S, Toyoshima S; Biol Pharm Bull 2001; 24(2): 119-122

The National CFIDS Foundation * 103 Aletha Rd, Needham Ma 02492 * (781) 449-3535 Fax (781) 449-8606